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心臟組織消化手冊

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Worthington組織消化指南

引言

組織消化酶主要用于組織培養(yǎng)和細胞生物學研究的原代細胞的分離和收集。盡管這些消化酶已經(jīng)普遍應用多年,但是對其組織消化和細胞收集的作用機制卻沒有全面的理解。  由此導致的結果便是選擇一種消化方法大多是隨機選擇或基于過去的經(jīng)驗,而不是在*理解消化原理的基礎上選擇的消化方案,以及如何根據(jù)具體實驗目的修正消化方案以獲得更好的實驗結果。

細胞分離程序的目的是zui大程度的獲得功能活性的單個細胞,而影響結果的原因有很多,包括(不限于)以下:

1、組織類型

2、物種來源

3、動物年齡

4、遺傳改造情況(如knockouts

5、所用的消化培養(yǎng)液

6、所用的酶

7、消化酶中的雜質

8、所用消化酶的工作濃度

9、溫度

10、孵育時間

通常前四個原因沒辦法選擇,而剩下的原因一般由經(jīng)驗決定以獲取zui適實驗結果。

通過查閱文獻來選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實驗數(shù)據(jù)搞得一頭霧水。這些巨大差異來源于組織胞外基質的復雜而動態(tài)的特性,以及過去常用的粗制、未提純的組織分離酶。另外,組織胞外基質由各種蛋白、糖蛋白、脂類和糖脂組成,這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育(年齡)之間存在明顯的差異,這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶,如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。

本指南總結了目前常用組織消化酶的zui適消化和細胞收集知識和方法,描述了標準的實驗步驟, 便于制定一個*的細胞分離方案,同時列舉了一些相關的參考文獻。

下表列出多種組織的適宜消化酶(含參考文獻,以組織類型和物種分類,詳見鏈接)

脂肪組織

腎上腺組織

骨組織

腦組織

軟骨組織

結腸組織

內皮組織

上皮組織

心臟

淋巴結

乳腺

其它組織

其他組織

肌組織

神經(jīng)組織

胰腺

腮腺

垂體

前列腺

生殖

動物鱗片

皮膚

脾臟

干細胞

胸腺

甲狀腺/甲狀旁腺

扁桃體

腫瘤

 

 

 

注:使用Worthington公司消化酶的文獻不限于上表鏈接中所列,一般而言,Worthington公司的組織消化酶可用于大多組織類型的消化。

 

心臟組織消化手冊

 

物種詳細來源細胞名稱消化用酶培養(yǎng)基參考文獻
牛 HeartTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and B綿羊 Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
(包括大鼠)Adrenal chromaffin 
 Paraneurons
CardiomyocytesCollagenase Type 2: 150 u/mlTyrode's solutionZhang,D., Wu, C., Qi, X., Meijering, R., Hoogstra-Berends, F., Tadevosyan, A., Cubukcuoglu, et al: Activation of Histone Deacetylase-6 Induces Contractile Dysfunction Through Derailment of alpha-tubulin Proteostasis in Experimental and Human Atrial Fibrillation., Circulation 129, 346-58, 2014
犬, adultAtrial and Ventricular MyocytesCollagenase Type 2: 0.065%See referenceBonilla, I., Sridhar, A., Nishijima, Y., Gyorke, S., Cardounel, A. and Carnes, C.: Differential Effects of the Peroxynitrite Donor, SIN-1, on Atrial and Ventricular Myocyte Electrophysiology., J Cardiovasc Pharmacol 61, 401-7, 2013
CardiomyocytesCollagenase Type 2: 300 u/mlTyrode's solutionXi, Y., Wu, G., Ai, T., Cheng, N., Kalisnik, J., Sun, J., Abbasi, S., Yang, D., Fan, C., Yuan, X., Wang, S. et al: Ionic Mechanisms Underlying the Effects of Vasoactive Intestinal Polypeptide on Canine Atrial Myocardium., Circ Arrhythm Electrophysiol 6, 976-83, 2013
Protease: 0.03%
Atrial MyocytesCollagenase Type 2: 0.03%Tyrode's solutionGan, T., Qiao, W., Xu, G., Zhou, X., Tang, B., Song, J., Li, Y., Zhang, J., Li, F., Mao, T. and Jiang, T.: Aging-Associated Changes in L-type Calcium Channels in the Left Atria of Dogs., Exp Ther Med 6, 919-924, 2013
CardiomyocytesCollagenase Type 2: 60 u/mlSee referenceHarleton, E., Besana, A., Comas, G., Danilo, P., Rosen, T., Argenziano, M., Rosen, M., Robinson, R. and Feinmark, S.: Ability to Induce Atrial Fibrillation in the Peri-Operative Period is Associated with Phosphorylation-Dependent Inhibition of TWIK Protein-Related Acid-Sensitive Potassium Channel 1 (TASK-1)., J Biol Chem 288, 2829-38, 2013
Protease XIV: 0.5 u/ml
Atrial and Ventricular MyocytesCollagenase Type 2: 0.05-0.08%See referenceZhang, H., Silva, J., Lin, Y., Verbsky, J., Lee, U., Kanter, E., Yamada, K., Schuessler, R. and Nichols, C.: Heterogeneity and Function of K(ATP) Channels in Canine Hearts., Heart Rhythm 10, 1576-83, 2013
犬, adultCardiomyocytesCollagenase Type 2: 0.05%HEPESCalloe, K., Nof, E., Jespersen, T., Di Diego, J., Chlus, N., Olesen, S.? Antzelevitch, C. and Cordeiro, J.: Comparison of the Effects of a Transient Outward Potassium Channel Activator on Currents Recorded from Atrial and Ventricular Cardiomyocytes., J Cardiovasc Electrophysiol 22, 1057-66, 2011
Protease Type XIV: 0.01%
犬, 20-25 kgCardiomyocytesCollagenase Type 2: 0.05%Tyrode's solutionBurashnikov, A., Zygmunt, A., Di Diego, J., Linhardt, G., Carlsson, L. and Antzelevitch, C.: AZD1305 Exerts Atrial Predominant Electrophysiological Actions and is Effective in Suppressing Atrial Fibrillation and Preventing its Reinduction in the Dog., J Cardiovasc Pharmacol 56, 80-90, 2010
犬, adultCardiomyocytesCollagenase Type 2: 0.05%M199Gavi, S., Yin, D., Shumay, E., Wang, H. and Malbon, C.: Insulin-Like Growth Factor-I Provokes Functional Antagonism and Internalization of Beta1-Adrenergic Receptors., Endocrinology 148, 2653-62, 2007
Protease: 0.008%
Atrial and Ventricular myocytesCollagenase Type 2: 0.065%Tyrode's solutionSchotten, U., de Haan, S., Verheule, S., Harks, E., Frechen, D., Bodewig, E., Greiser, M., Ram, R., Maessen, J., Kelm, M., Allessie, M. and Van Wagoner, D.: Blockade of Atrial-Specific K+-currents Increases Atrial but not Ventricular Contractility by Enhancing Reverse Mode Na+/Ca2+-Exchange.,Cardiovasc Res 73, 37-47, 2007
Atrial myocytesCollagenase Type 2: 0.013%HEPESBaba, S., Dun, W., Hirose, M. and Boyden, P.: Sodium Current Function in Adult and Aged Canine Atrial Cells., Am J Physiol Heart Circ Physiol 291, H756-61, 2006
犬, 20-30 kgCardiomyocytesCollagenase Type 2: 110 u/mlTyrode's solutionSun, H., Chartier, D., Leblanc, N. and Nat, S.: Intracellular Calcium Changes and Tachycardia-Induced Contractile Dysfunction in Canine Atrial Myocytes., Cardiovasc Res 49, 751-61, 2001
Ventricular and Atrial MyocytesCollagenase Type 4: 0.04%HEPES/Tyrode'sGintant, G.: Two Components of Delayed Rectifier Current in Canine Atrium and Ventricle. Does IKs Play a Role in the Reverse Rate Dependence of Class III Agents?, Circ Res 78, 26-37, 1996
Collagenase Type 2: 125 u/ml
Dog, mongrel, adultMyocytesHyaluronidase: 0.1%Joklik's MEMSpanier, A. and Weglicki, W.: Ca2+-Tolerant Adult Canine Myocytes: Preparation and Response to Anoxia/Acidosis, Am J Physiol 243, H448, 1982
雞, embryonicCardiomyocytesTrypsin: 0.13%HBSSBlech-Hermoni, Y. and Ladd, A.: Identification of Transcripts Regulated by CUG-BP, Elav-like Family Member 1 (CELF1) in Primary Embryonic Cardiomyocytes by RNA-seq., Genom Data 6, 74-76, 2015
Collagenase Type 2: 0.13%
Deoxyribonuclease I: 0.033%
雞 embryos, 9-11 day incubatedCardiomyocytesTrypsin: 0.25%PBSEschenhagen, T., Fink, C., Remmers, U., Scholz, H., Wattchow, J., Weil, J., et al.: Three-Dimensional Reconstitution of Embryonic Cardiomyocytes in a Collagen Matrix: A New Heart Muscle Model System, FASEB J 11, 683, 1997
小雞, embryos, 10-11 dayCardiomyocytesTrypsin: 0.17%HBSS, CMFWang, S., Greaser, M.L., Schultz, E., Bulinski, J.C., Lin, J.J., and Lessard, J.L.: Studies on Cardiac Myofibrillogenesis With Antibodies to Titin, Actin, Tropomyosin, and Myosin, J Cell Biol 107, 1075, 1988
小雞, 10 dayVentricularTrypsin: 0.025%HBSS, CMFKim, D., Okada, A., and Smith, T.W.: Control of Cytosolic Calcium Activity During Low Sodium Exposure in Cultured Chick Heart Cells, Circ Res 61, 29, 1987
小雞, embryo, 11 dayCardiomyocytesTrypsin: 0.05%HBSS, CMFJacob, R., Lieberman, M., Murphy, E., and Piwnica-Worms, D.: Effect of Sodium-Potassium Pump Inhibition and Low Sodium on Membrane Potential in Cultured Embryonic Chick Heart Cells, J Physiol 387, 549, 1987
小雞, embryonic, 11-12-day-oldMyocytesTrypsin: 0.025%CMF solutionMurphy, E., Aiton, J., Russell, C., and Lieberman, M.: Calcium Elevation in Cultured Heart Cells: Its Role in Cell Injury, Am J Physiol Cell Physiol 245 (14), C316, 1983
小雞, 8 day embryo MyocytesTrypsin NF 1:250: 0.25%Rinaldini's buffer solution, CFGross, W., Schopf-Ebner, E., and Bucher, O.: Technique for the Preparation of Homogeneous Cultures of Isolated Heart Muscle Cells, Exp Cell Res 53, 1, 1968
(also rat, neonate and mouse)
Hen, White LeghornHeartTrypsin: 0.025%Medium 199Dehann, R.: Regulation of Spontaneous Activity and Growth of Embryonic Chick Heart Cells in Tissue Culture, Dev Biol 16, 216, 1967
小雞embryos, 5 dayHeartTrypsin: 3.0%Tyrode's solution, CMFSteinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res 30, 257, 1963
Liver
貓科動物貓, mongrel, adult, 1.8-2.8 KgMyocytesCollagenase Type 2: 0.12%Kreb's Henseleit, CFSilver, L., Hemwall, E., Marino, T., and Houser, S.: Isolation and Morphology of Calcium-Tolerant Feline Ventricular Myocytes, Am J Physiol 245, H891, 1983
Ventricular
魚, 斑馬魚CardiomyocytesCollagenase Type 2: 0.5%MEMSander, V., Sune, G., Jopling, C., Morera, C. and Belmonte J.: Isolation and In Vitro Culture of Primary Cardiomyocytes from Adult Zebrafish Hearts., Nat Protoc 8, 800-9, 2013
Collagenase Type 4: 0.5%
青蛙青蛙 (Rana esculenta)MyocytesTrypsin: 0.04%CF RingerArrio-Dupont, M., and de Nay, D.: High Yield Preparation of Calcium-Tolerant Myocytes From Frog Ventricles, Biol Cell 54, 164, 1985
豚鼠Guinea pig, adultCardiomyocytesCollagenase Type 2: 100 u/mlM-199Zorn-Pauly K, Schaffer P, Pelzmann B, Bernhart E, Lang P, and Koidl B: L-type and T-type Ca2+ current in cultured ventricular guinea pig myocytes,Physiol Res 53(4), 369, 2004
豚鼠豚鼠CardiomyocytesCollagenase Type 2: 100-200 u/mlSee referenceDhamoon, A, Pandit, S., Sarmast, F., Parisian, K., Guha, P., Li, Y., Bagwe, S., Taffet, S. and Anumonwo, J.: Unique Kir2.x Properties Determine Regional and Species Differences in the Cardiac Inward Rectifier K+ Current.,Circ Res 94, 1332-9, 2004
豚鼠Guinea pig, female, 300-350gCoronary endothelialCollagenase Type 2: 0.1%see referenceBuxton I L, Kaiser R A, Oxhorn B C, Cheek D J: Evidence supporting the Nucleotide Axis Hypothesis: ATP release and metabolism by coronary endothelium, Am J Physiol Heart Circ Physiol 281, H1657-66, 2001
豚鼠Guinea pig, 250-450 gEndothelialCollagenase Type 2: 0.15%Perfusing solutionPreisig-Muller, R., Mederos Y Schnitzler, M., Derst, C., and Daut J.:Separation of Cardiomyocytes and Coronary Endothelial Cells for Cell-Specific RT-PCR, Am J Physiol 277, H413, 1999
(see reference)
豚鼠豚鼠, male, 450-600 gMyocytesHyaluronidase: 0.02%Krebs Henseleit bicarbonate bufferStemmer, P., Akera,T., Brody, T., Rardon, D., and Watanabe, A.: Isolation and Enrichment of Ca2+-Tolerant Myocytes for Biochemical Experiments from Guinea-Pig Heart, Life Sci 44, 1231, 1989
豚鼠豚鼠CardiomyocytesCollagenase: 0.04%Tyrode solution, CFIshihara, K., Mitsuiye, T., Noma, A., and Takano, M.: The Mg2+ Block and Intrinsic Gating Underlying Inward Rectification of the K+ Current in Guinea-Pig Cardiac Myocytes, J Physiol 419, 297, 1989
豚鼠豚鼠MyocytesHyaluronidase: 0.10%Bicarbonate buffer, CFBridge, J., Spitzer, K., and Ershler, P.: Relaxation of Isolated Ventricular Cardiomyocytes by a Voltage-Dependent, Science 241, 823, 1988
人類人類CardiacCollagenase: 0.045%DPBSHolt-Casper, D., Theisen, J., Moreno, A., Warren, M., Silva, F., Grainger, D., Bull, D. and Pa, A.: Novel Xeno-free Human Heart Matrix-Derived Three-Dimensional Scaffolds., J Transl Med 13, 194, 2015
Pancreatin: 0.1%
人類人類Cardiac stemCollagenase Type 2: 0.25%Joklik modified Eagle抯Avolio, E., Gianfranceschi, G., Cesselli, D., Caragnano, A., Athanasakis, E., Katare, R., Meloni, M., Palma, A., Barchiesi, A., Vascotto, C., Toffoletto, B., Mazzega, E., Finato, N., Aresu, G. and Livi, U.: Ex vivo Molecular Rejuvenation Improves the Therapeutic Activity of Senescent Human Cardiac Stem Cells in a Mouse Model of Myocardial Infarction., Stem Cells 32, 2373, 2014
人類人類Atrial myocytesCollagenase Type 1: 286 u/mlsee referencesVoigt, N., Zhou, X. and Dobrev, D.: Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents., J Vis Exp , e50235, 2013
Protease: 5 u/ml
人類人類MyofibersCollagenase Type 1: 0.3%see referenceAnderson, E., Rodriguez, E., Anderson, C., Thayne, K., Chitwood, W. and Kypson, A.: Increased Propensity for Cell Death in Diabetic Human Heart is Mediated by Mitochondrial-Dependent Pathways., Am J Physiol Heart Circ Physiol 300, H118, 2011
人類人類Atrial cardiomyocytesCollagenase Type 1: 286 u/mlsee referenceVoigt, N., Makary, S., Nat, S. and Dobrev, D.: Voltage-Clamp-Based Methods for the Detection of Constitutively Active Acetylcholine-Gated I(K,ACh) Channels in the Diseased Heart., Methods Enzymol 484, 653, 2010
Protease: 5 u/ml
人類人類Coronary artery smooth muscleCollagenase Type 2: 0.1%DMEMJensen, B., Swigart, P., Laden, M., DeMarco, T., Hoopes, C. and Simpson, P.: The Alpha-1D Is the Predominant Alpha-1-Adrenergic Receptor Subtype in Human Epicardial Coronary Arteries., J Am Coll Cardiol 54, 1137, 2009
Elastase: 0.05%
人類人類Smooth muscle aorticCollagenase Type 1: 0.2%DMEMMathew, S., Tustison, K., Sugatani, T., Chaudhary, L., Rifas, L. and Hruska, K.: The Mechanism of Phosphorus as a Cardiovascular Risk Factor in CKD., J Am Soc Nephrol 19, 1092, 2008
Collagenase Type 2: 0.1%
Elastase: 0.025%
人類人類 (n=16, age: 60 &plusmin;3 years)MyocardialHyaluronidase: 0.05%Medium 199Mukerjee, R., Multani, M., Sample, J., Dowdy, K., Zellner, J., Hoover, D., and Spinale, F.: Effects of adrenomedullin on human myocyte contractile function and beta-adrenergic response, J Cardiovasc Pharmacol 7 (4), 235, 2002
人類人類CardiomyocytesCollagenase Type 2: 0.05%HEPES solutionTodor Anastassia, Sharov Victor G, Tanhehco Elaine J, Silverman Norman, Bernabei Alvise, Sabbah Hani N: Hypoxia-induced cleavage of caspase-3 and DFF45/ICAD in human failed cardiomyocytes, Am J Physiol Heart Circ Physiol 283, H990-5, 2002
Collagenase Type 1: 0.025%
Protease XIV: 0.013%
人類人類HeartCollagenase Type 2: 200 u/mlTyrode's solutionHoppe, U., Jansen, E., Sudkamp, M., and Beukelmann, D.:Hyperpolarization-Activated Inward Current in Ventricular Myocytes From Normal and Failing Human Hearts, Circulation 97, 55, 1998
Myocytes
人類人類Atrial myocytesCollagenase Type 2: 0.1%see referenceVan Wagoner, D., Pond, A., McCarthy, P., Trimmer, J. and Nerbonne, J.:Outward K+ Current Densities and Kv1.5 Expression are Reduced in Chronic Human Atrial Fibrillation., Circ Res 80, 772-81, 1997
Protease Type XIV: 0.04%
人類人類, fetal, 14.5 week gestationMyocytesHyaluronidase: 0.1%HBSS with CalciumGoldman, B., and Wurzel, J.: Effects of Subc*tion and Culture Medium on Differentiation of Human Fetal Cardiac Myocytes, In Vitro Cell Dev Biol 28, 109, 1992
人類人類, both sexes, 18-81 yearsMyocardialCollagenase: 0.14%HBSS, CMFSmith, D., Glover, J., Townsend, L., and Maupin, E.: A Method for the Harvest, Culture, and Characterization of Human Adult Atrial Myocardial Cells: Correlation with Age of Donor, In Vitro Cell Dev Biol 27, 914, 1991
Atrial
人類人類, adultMyocytesTrypsin: 0.25%Joklik's MEMBugaisky, L.B.: Biology of Isolated Adult Cardiac Myocytes,Isolation and Culture of Human Adult Cardiac Myocytes,(Clark,Decker,Borg,eds), Elsevier Science Publishing Co., Inc., , 1988
人類人類Thoracic aortaCollagenase: 0.2%Phosphate buffer w/NaClHassler, O., Wiren, M., and Herbertsson, S.: The Elastic Coat of the Arterial Wall Studied with the Aid of Collagenase, Acta Pathol Microbiol Scand 57, 15, 1963
無脊椎動物E. cirhosa章魚, 260-352 克Systemic heart cardiomyocytesCollagenase: 0.025%see referenceAltimiras J., Hove-Madsen L., and Gesser H.: Ca(2+) Uptake in the Sarcoplasmic Reticulum from the Systemic Heart of Octopod Cephalopods, J Exp Biol 202, 2531, 1999
Trypsin: 0.02%
小鼠小鼠, 15-30 gmVentricular myocytesCollagenase Type 2: 0.12%see referenceHenn, M., Janjua, M., Kanter, E., Makepeace, C., Schuessler, R., Nichols, C. and Lawton, J.: Adenosine Triphosphate-Sensitive Potassium Channel Kir Subunits Implicated in Cardioprotection by Diazoxide., J Am Heart Assoc 4, , 2015
小鼠小鼠, 3-4 weekCardiac progenitorCollagenase Type 1: 0.1%DMEMWang, H., Chen, H., Feng, B., Wang, X., He, X., Hu, R., Yin, M., Wang, W., Fu, W. and Xu, Z.: Isolation and Characterization of a Sca-1+/CD31- Progenitor Cell Lineage Derived From Mouse Heart Tissue., BMC Biotech4, 75, 2014
小鼠小鼠, adultCardiac ProgenitorCollagenase Type 2: 600 u/mlMEMSantos Nascimento, D, Mosqueira, D, Sousa, L, Teixeira, M, Filipe, M, Resende, T, Araujo, A, Valente, M, Almeida, J, Martins, J, Santos, J, Barcia, R, Cruz, P, Cruz, H and Pinto-d-O, P: Human Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Attenuate Remodeling after Myocardial Infarction by Proangiogenic, Antiapoptotic, and Endogenous Cell-Activation Mechanisms., Stem Cell Res Ther 5, 5, 2014
Deoxyribonuclease I: 60 u/ml
小鼠小鼠Cardiac progenitorCollagenase Type 2: 0.1%HBSSZafiriou, M., Noack, C., Unsold, B., Didie, M., Pavlova, E., Fischer, H., Reichardt, H., Bergmann, M., El-Armouche, A., Zimmermann, W., Zelarayan, L.: Erythropoietin Responsive Cardiomyogenic Cells Contribute to Heart Repair Post Myocardial Infarction., Stem Cells 32, 2480, 2014
Neutral Protease: 2.4 u/ml
小鼠小鼠, adultCardiomyocytesCollagenase Type 2: 0.08%see referenceKohncke, C. Lisewski, U., Schleussner, L., Gaertner, C., Reichert, S. and Roepke, T.: Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes., J Vis Exp 73, e50145, 2013
小鼠小鼠CardiomyocytesCollagenase Type 2: 150 u/mlMEMHennessey, J., Wei, E. and Pitt, G.: Fibroblast Growth Factor Homologous Factors Modulate Cardiac Calcium Channels., Circ Res 113, 381-8, 2013
小鼠小鼠CardiomyocytesCollagenase Type 2: 360 u/mlL-15Touchberry, C., Green, T., Tchikrizov, V., Mannix, J., Mao, T., Carney, B., Girgis, M., Vincent, R., Wetmore, L., Dawn, B., Bonewald, L., Stubbs, J. and Wacker, M.: FGF23 is a Novel Regulator of Intracellular Calcium and Cardiac Contractility in Addition to Cardiac Hypertrophy., Am J Physiol Endocrinol Metab 304, E863, 2013
Papain: 0.4 u/ml
Deoxyribonuclease I: 40 u/ml
小鼠小鼠, 2-6 monthSinoatrial nodeCollagenase Type 2: 229 u/mlTyrode's solutionChris, C., Cardona, N., Mesirca, P., Herrmann, S., Hofmann, F., Striessnig, J., Ludwig, A., Mangoni, M. and Lee, A.: Distinct Localization and Modulation of Cav1.2 and Cav1.3 L-type Ca2+ Channels in Mouse Sinoatrial Node., J Physiol 590, 6327, 2012
Elastase: 1.9 u/ml
Protease Type XIV: 0.9 u/ml
小鼠小鼠, male, 8-25 weekCardiomyocytesCollagenase Type 2: 59 u/mlMEMCarley, A. and Kleinfeld, A.: Fatty Acid (FFA) Transport in Cardiomyocytes Revealed by Imaging Unbound FFA is Mediated by an FFA Pump Modulated by the CD36 Protein., J Biol Chem 286, 4589-97, 2011
小鼠小鼠Aortic adventitial leukocytesCollagenase Type 2: 300 u/mlPBSButcher, M., Herre, M., Ley, K. and Galkina, E.: Flow Cytometry Analysis of Immune Cells Within Murine Aortas., J Vis Exp 53, e2828, 2011
Elastase: 5.6 u/ml
小鼠小鼠, adultCardiomyocytesCollagenase Type 1: 0.1-0.4%see referenceFlynn, J., Santana, L. and Melov, S.: Single Cell Transcriptional Profiling of Adult Mouse Cardiomyocytes., J Vis Exp 58, e3302, 2011
Protease Type XIV: 0.004-.02%
小鼠小鼠, male, 6 weekCoronary endothelial cellsCollagenase Type 2: 0.1%M199Makino, A., Suarez, J., Wang, H., Belke, D., Scott, B. and Dillmann, W.:Thyroid Hormone Receptor-beta is Associated With Coronary Angiogenesis During Pathological Cardiac Hypertrophy., Endocrinology 150, 2008, 2009
Neutral Protease: 0.6 u/ml
小鼠小鼠, adultVentricular myoctesCollagenase: 0.1%M199Zhang, Y, Kanter, E., Laing, J., Aprhys, C., Johns, D., Kardami, E. and Yamada, K.: Connexin43 Expression Levels Influence Intercellular Coupling and Cell Proliferation of Native Murine Cardiac Fibroblasts., Cell Commun Adhes 15, 289, 2008
小鼠小鼠, adult, 9-13 weekCardiomyocytesCollagenase Type 2: 620 u/mlMyocyte buffer see referenceKabaeva, Z., Zhao, M. and Michele, D.: Blebbistatin Extends Culture Life of Adult Mouse Cardiac Myocytes and Allows Efficient and Stable Transgene Expression., Am J Physiol Heart Circ Physiol 294, H1667-74, 2008
Protease XIV: 0.104 u/ml
Deoxyribonuclease I: 0.0015%
小鼠小鼠, adultCardiomyocytesCollagenase: see referenceHBSSO'Connell Timothy D, Rodrigo Manoj C, Simpson Paul C: Isolation and culture of adult mouse cardiac myocytes, Methods Mol Biol 357, 271-96, 2007
小鼠小鼠Smooth muscle cellsCollagenase Type 2: 300 u/mlF10 HamVerheye, S., Martinet, W., Kockx, M., Knaapen, M., Salu, K., Timmermans, J., Ellis, J., Kilpatrick, D., DeMayer, G.: Selective Clearance of Macrophages in Atherosclerotic Plaques by Autophagy, J Am Coll Cardiol49, 706, 2007
Elastase: 5 u/ml
小鼠小鼠, adult, 6-12 weekCardiomyocytesCollagenase Type 2: 0.1%see referenceShioya, T: A Simple Technique for Isolating Healthy Heart Cells from Mouse Models., J Physiol Sci 57, 327-35, 2007
Trypsin: 0.006%
Protease XIV: 0.006%
小鼠小鼠Ventricular myocytes and mesenteric arterial SMCCollagenase Type 2: 0.06%Krebs-RingerLu Tong, Ye Dan, Wang Xiaoli, Seubert John M, Graves Joan P, Bradbury J Alyce, Zeldin Darryl C, Lee Hon-Chi: Cardiac and vascular KATP channels in rats are activated by endogenous epoxyeicosatrienoic acids through different mechanisms, J Physiol 575, 627-44, 2006
Papain: 0.175%
小鼠小鼠CardiomyocytesCollagenase Type 1: 0.17%PBSChen, H., Yong, W., Ren, S., Shen, W., He, Y., Cox, K., Zhu, W., Li, W., Soonpaa, M., Payne, RM, Franco, D., Field, L., Rosen, V., Wang, Y. and Shou, W.: Overexpression of Bone Morphogenetic Protein 10 in Myocardium Disrupts Cardiac Postnatal Hypertrophic Growth., J Biol Chem 281, 27481, 2006
小鼠小鼠, neonatalCardiomyoctesCollagenase Type 1: 150 u/mlM199Takahashi N, Wang X, Tanabe S, Uramoto H, Jishage K, Uchida S, Sasaki S, Okada Y.: ClC-3-independent sensitivity of apoptosis to Cl- channel blockers in mouse cardiomyocytes, Cell Physiol Biochem 15, 263, 2005
Trypsin: 0.01%
小鼠小鼠, 1 dayCardiomyoctesNCIS kit: per instructionsEBSSPotts, Malia B., Vaughn, Allyson E., McDonough, Holly, Patterson, Cam, and Deshmukh, Mohanish: Reduced Apaf-1 levels in cardiomyocytes engage strict regulation of apoptosis by endogenous XIAP, J Cell Biol 171, 925, 2005
小鼠小鼠Aortic endothelialCollagenase Type 2: 0.2%DMEMKobayashi, M., Inoue, K., Warabi, E., Minami, T. and Kodama, T.: A Simple Method of Isolating Mouse Aortic Endothelial Cells., J Atheroscler Thromb 12, 138, 2005
小鼠小鼠Smooth muscle cellsCollagenase Type 1: 0.1%DMEMFukumoto Yoshihiro, Deguchi Jun-o, Libby Peter, Rabkin-Aikawa Elena, Sakata Yasuhiko, Chin Michael T, Hill Christopher C, Lawler Patrick R, Varo Nerea, Schoen Frederick J, Krane Stephen M, Aikawa Masanori:Genetically determined resistance to collagenase action augments interstitial collagen accumulation in atherosclerotic plaques, Circulation 110, 1953-9, 2004
Elastase: 0.0125%
小鼠小鼠, female, 14-18 gCardiomyocytesCollagenase Type 2: 0.02%see referenceZhang Sui, Wang Dachun, Estrov Zeev, Raj Sean, Willerson James T, Yeh Edward T H: Both cell fusion and transdifferentiation account for the transformation of human peripheral blood CD34-positive cells into cardiomyocytes in vivo, Circulation 110, 3803-7, 2004
Elastase: 0.03%
Pancreatin: 0.06%
小鼠小鼠, 6-8 weekVascular smooth musclePapain: 10 u/mlEBSSQian, Q., Hunter, L., Li, M., Marin-Padilla, M., Prakash, Y., Somlo, S., Harris, P., Torres, V., and Sieck, G.: Pkd2 Haploinsufficiency Alters Intracellular Calcium Regulation in Vascular Smooth Muscle Cells, Hum Mol Genet 12(15), 1875, 2003
Elastase: .005%
Collagenase: 0.05%
Deoxyribonuclease I: 1000 u/ml
PDS kit: with modifications
小鼠小鼠CardiomyocytesCollagenase Type 2: 0.2%DMEM/F-12Rybkin, I., Markham, D., Yan, Z., Bassel-Duby, R., Williams, R., and Olson, E.: Conditional Expression of SV40 T-antigen in Mouse Cardiomyocytes Facilitates an Inducible Switch from Proliferation to Differentiation, J Biol Chem 278(18), 15927, 2003
Pancreatin: 0.06%
小鼠小鼠, neonatal 2-4 dayAtrial myocytesNCIS kit: with modifications see referenceL-15Bettahi, I,, Marker, C., Roman, M., and Wickman, K.: Contribution of the Kir3.1 Subunit to the Muscarinic-gated Atrial Potassium Channel IKACh, J Biol Chem 277, 48282, 2002
小鼠小鼠, neonatal and adultCardiomyocytesCollagenase Type 2: 0.03-0.2%Joklik's MEMSchreiber, K., Paquet, L., Allen, B., and Rindt, H.: Protein Kinase C Isoform Expression and Activity in the Mouse Heart, Am J Physiol Heart Circ Physiol281(5), H2062, 2001
小鼠小鼠, NTg and Tg, 8 - 12 weekMyocytesCollagenase Type 2: 150 u/mlJoklik's MEMNelson, D., Setser, E., Hall, D., Schwartz, S., Hewitt, T., Klevitsky, R., Osinska, H., Bellgrau, D., Duke, R., and Robbins, J.: Proinflammatory Consequences of Transgenic Fas Ligand Expression in the Heart, J Clin Invest 105 (9), 1199, 2000
小鼠小鼠, 1-6 dayCardiomyocytesCollagenase Type 2: 0.1%PBSWatzka SB, Lucien J, Shimada M, Edwards V, Yeger H, Hannigan G, and Coles JG.: Selection of viable cardiomyocytes for cell transplantation using three-dimensional tissue culture, Transplantation 70, 1310, 2000
Deoxyribonuclease I: 0.002%
小鼠小鼠, 2-4 moVentricular myocytesCollagenase Type 2: 0.05%MEMZhou YY, Wang SQ, Zhu WZ, Chruscinski A, Kobilka BK, Ziman B, Wang S, Lakatta EG, Cheng H, Xiao RP: Culture and adenoviral infection of adult mouse cardiac myocytes: methods for cellular genetic physiology, Am J Physiol Heart Circ Physiol 279, H429-36, 2000
Collagenase Type 4: 0.05%
Protease XIV: 0.002%
小鼠小鼠CardiomyocytesCollagenase Type 2: 150 u/mlJoklik's MEMChristensen G, Minamisawa S, Gruber PJ, Wang Y, Chien KR: High-efficiency, long-term cardiac expression of foreign genes in living mouse embryos and neonates, Circulation 101, 178-84, 2000
小鼠小鼠, 1-3 dayCardiomyocytesTrypsin: 0.25%FBS-MEMWang, G., and Kang, Y.: Inhibition of Doxorubicin Toxicity in Cultured Neonatal Mouse Cardiomyocytes with Elevated Metallothionein Levels, J Pharmacol Exp Ther 288(3), 938, 1999
小鼠小鼠, 1 dayCardiomyocytesNCIS kit: with modifications see referenceL-15Lader, A., Kwiatkowski, D., and Cantiello, H.: Role of Gelsolin in the Actin Filament Regulation of Cardiac L-type Calcium Channels, Am J Physiol 277, C1277, 1999
小鼠小鼠, 3-7 monthFibroblasts, mesangial, smooth muscleTrypsin: 0.25%DMEMBradshaw AD, Francki A, Motamed K, Howe C, Sage EH: Primary mesenchymal cells isolated from SPARC-null mice exhibit altered morphology and rates of proliferation, Mol Biol Cell 10, 1569-79, 1999
Collagenase: see reference
Soybean Trypsin Inhibitor: .05%
小鼠小鼠 (BALB/c), adultMyocytesCollagenase Type 2: 0.04%Tyrode's solutionFelzen, B., Shilkrut, M., Less, H., Sarapov, I., Maor, G., Coleman, R., Robinson, R., et al.: Fas (CD95/Apo-1)-Mediated Damage to Ventricular Myocytes Induced by Cytotoxic T Lymphocytes from Perforin-Deficient Mice,Circ Res 82, 438, 1998
小鼠小鼠, 1 dayCardiomyocytesNCIS kit: with modifications see referenceL-15Lader, A., Xiao, Y., Ishikawa, Y., Cui, Y., Vatner, D., Vatner, S., Homcy, C., Cantiello, H.: Cardiac Gsalpha Overexpression Enhances L-type Calcium Channels Through an Adenylyl Cyclase Independent Pathway, Proc Natl Acad Sci U S A 95, 9669, 1998
小鼠小鼠, P0-P1CardiomyocytesCollagenase: 0.045%DMEM, Medium 199Arber, S., Hunter, J., Ross, J., Hongo, M., Sansig, G., Borg, J., Perriard, J., Chien, K., and Caroni, P.: MLP-Deficient Mice Exhibit a Disruption of Cardiac Cytoarchitectural Organization, Dilated Cadiomyopathy, and Heart Failure,Cell 88, 393, 1997
小鼠小鼠, 6-9 or 21-41 dayVentricular and atrial myocytesCollagenase Type 2: 95 u/mlDMEM/TyrodesValenzuela, D., Han, X., Mende, U., Fankhauser, C., Mashimo, H., Huang, P., Pfeffer, J., Neer, E., and Fishman, M.: G alpha(o) is Necessary for Muscarinic Regulation of Ca2+ Channels in Mouse Heart, Proc Natl Acad Sci U S A 94(5), 1727, 1997
Hyaluronidase: 172.5 u/ml
Trypsin: 0.002%
Deoxyribonuclease I: 60u/ml
小鼠小鼠CardiomyocyteCollagenase: 0.17 %PBSSoonpaa, M., Kim, K., Pajak, aL., Franklin, M., and Field, L.:Cardiomyocyte DNA Synthesis and Binucleation During Murine Development,Am J Physiol 271, H2183, 1996
小鼠小鼠 embryosMyocytesCollagenase Type 1: 0.1%DMEMWobus, A., Kleppisch, T., Maltsev, V., and Hescheler, J.: Cardiomyocyte-Like Cells Differentiated In Vitro From Embryonic Carcinoma Cells P19 are Characterized by Functional Expression of Adrenoceptors and Ca2+ Channels,In Vitro Cell Dev Biol 30A, 425, 1994
小鼠小鼠 MyocytesTrypsin NF 1:250: 0.25%Rinaldini's buffer solution, CFGross, W., Schopf-Ebner, E., and Bucher, O.: Technique for the Preparation of Homogeneous Cultures of Isolated Heart Muscle Cells, Exp Cell Res 53, 1, 1968
(also rat, neonate and chick, 8 day embryo)
綿羊綿羊Atrial myocytesCollagenase Type 2: 0.055%Tyrode's solutionVoigt, N., Pearman, C., Dobrev, D. and Dibb, K.: Methods for Isolating Atrial Cells from Large Mammals and Humans., J Mol Cell Cardiol 86, 187-98, 2015
Protease XIV: 0.006%
綿羊綿羊CardiomyocytesCollagenase Type 2: 0.055%See referenceDibb, K., Clarke, J., Horn, M., Richards, M., Graham, H., Eisner, D. and Trafford, A.: Characterization of an Extensive Transverse Tubular Network in Sheep Atrial Myocytes and its Depletion in Heart Failure., Circ Heart Fail 2, 482-9, 2009
Protease Type XIV: 0.006%
綿羊綿羊CardiomyocytesCollagenase Type 2: 100-200 u/mlSee referenceDhamoon, A, Pandit, S., Sarmast, F., Parisian, K., Guha, P., Li, Y., Bagwe, S., Taffet, S. and Anumonwo, J.: Unique Kir2.x Properties Determine Regional and Species Differences in the Cardiac Inward Rectifier K+ Current.,Circ Res 94, 1332-9, 2004
Aortic smooth muscle and intersitial cellsCollagenase: 600 u/mlPBSButcher, J. and Nerem, R.: Porcine Aortic Valve Interstitial Cells in Three-Dimensional Culture: Comparison of Phenotype with Aortic Smooth Muscle Cells., J Heart Valve Dis 13, 478, 2004
Coronary vascular smooth muscle (PCVSMCs)Trypsin: 0.037%HEPESChrist, M., Gunther, A., Heck, M., Schmidt, B., Falkenstein, E., and Wehling, M.: Aldosterone, Not Estradiol, Is the Physiological Agonist for Rapid Increases in cAMP in Vascular Smooth Muscle Cells, Circulation 99, 1485, 1999
Miniature swine, adult, female, 25-40 kg, familiarized with treadmill exerciseCoronary smooth muscleTrypsin: 0.1%Krebs bicarbonate solutionBowles, D., Hu, Q., Laughlin, M., and Sturek, M.: Exercise Training Increases L-type Calcium Current Density in Coronary Smooth Muscle, Am J Physiol 275 (44), H2159, 1998
兔, New Zealand, 2.3-2.9 kgCardiomyocytesCollagenase Type 1: 0.05%Krebs-HenseleitFarkas, A., Acsai, K., Nagy, N., Toth, A., Fulop, F., Seprenyi, G., Birinyi, P., Nanasi, P., Forster, T., Csanady, M., Papp, J., Varrro, A. and Farkas, A.:Na(+)/Ca(2+) Exchanger Inhibition Exerts a Positive Inotropic Effect in the Rat Heart, but Fails to Influence the Contractility of the Rabbit Heart., Br J Pharmacol 154, 93, 2008
Hyaluronidase: see reference
Protease: see reference
Smooth muscle cellsCollagenase Type 2: 300 u/mlF10 HamVerheye, S., Martinet, W., Kockx, M., Knaapen, M., Salu, K., Timmermans, J., Ellis, J., Kilpatrick, D., DeMayer, G.: Selective Clearance of Macrophages in Atherosclerotic Plaques by Autophagy, J Am Coll Cardiol49, 706, 2007
Elastase: 5 u/ml
兔, New Zealand, 2 kgCardiomyocytes and fibroblastsCollagenase: 0.06%Medium 199Driesen Ronald B, Dispersyn Gerrit D, Verheyen Fons K, van den Eijnde Stefan M, Hofstra Leo, Thone Fred, Dijkstra Petra, Debie Wiel, Borgers Marcel, Ramaekers Frans C S: Partial cell fusion: a newly recognized type of communication between dedifferentiating cardiomyocytes and fibroblasts,Cardiovasc Res 68, 37-46, 2005
兔, New Zealand White, 2-3 kgMyocytesCollagenase Type 2: 0.1%HEPESSpitzer, K., Ershler, P., Skolnick, R., and Vaughan-Jones, R.: Generation of Intracellular pH Gradients in Single Cardiac Myocytes with a Microperfusion System, Am J Physiol 278, H1371, 2000
兔, New Zealand, white, either sexMyocytesCollagenase Type 2: 0.05%EGTA-KBSedarat, F., Xu, L., Moore, E., and Tibbits, G.: Colocalization of Dihydropyridine and Ryanodine Receptors in Neonate Rabbit Heart Using Confocal Microscopy, Am J Physiol Heart Circ Physiol 279, H202, 2000
兔, adult 1.5-2.5 kgTricuspid valve cellsCollagenase: 0.8 mg/mlHEPES Tyrode solutionAnumonwo JM, Delmar M, and Jalife J.: Electrophysiology of single heart cells from the rabbit tricuspid valve, J Physiol 425, 145, 1990
兔, New Zealand, white, male, 2-3 kgCardiomyocytesHyaluronidase: 0.5%Eagle's MEMBuxton, I., and Brunton, L.: Compartments of Cyclic AMP and Protein Kinase in Mammalian Cardiomyocytes, J Biol Chem 258 (17), 10233, 1983
兔, adult, 2 kgMyocytesHyaluronidase: 0.007%Kreb's RingerDani, A., Cittadini, A., Flamini, G., Festuccia, G., and Terranova, T.:Preparation and Some Properties of Isolated Beating Myocytes from Adult Rabbit Heart, J Mol Cell Cardiol 9, 777, 1977
大鼠大鼠, 1-2 dayCardiomyocytesNCIS kit: per instructionsMEMXie, L., Pi, X., Townley-Tilson, WH., Li, N., Wehrens, X., Entman, M., Taffet, G., Mishra, A., Peng, J., Schisler, J., Meissner, G. and Patterson, C.: PHD2/3-dependent Hydroxylation Tunes Cardiac Response to 尾-adrenergic Stress via Phospholamban., J Clin Invest 125, 2759-71, 2015
大鼠大鼠, Wistar, 250-350gVentricular myocytesCollagenase Type 2: 251 u/mlTyrode solutionCalderon-Sachez, E., Domi璶guez-Rodriguez, A., Lopez-Haldon, J., Jimenez-Navarro, M., Gomez, A., Smani, T. and Ordonez, A.:Cardioprotective Effect of Ranolazine in the Process of Ischemia-reperfusion in Adult Rat Cardiomyocytes., Rev Esp Cardiol (Engl Ed) , , 2015
大鼠大鼠, SD, 8-10 weekVentricular myocytesCollagenase: 0.08%Tyrode's SolutionLee, J., Ha, J. and Leem, C.: A Novel Nicotinamide Adenine Dinucleotide Correction Method for Mitochondrial Ca(2+) Measurement with FURA-2-FF in Single Permeabilized Ventricular Myocytes of Rat., Korean J Physiol Pharmacol 19, 373-82, 2015
Pronase: 0.004%
大鼠大鼠, SD, 1 dayCardiomyocytesTrypsin: 0.025%HBSSClark, A. and Naya, F.: MicroRNAs in the Myocyte Enhancer Factor 2 (MEF2)-regulated Gtl2-Dio3 Noncoding RNA Locus Promote Cardiomyocyte Proliferation by Targeting the Transcriptional Coactivator Cited2., J Biol Chem 290, 23162-72, 2015
Collagenase Type 2: 1.0%
大鼠大鼠, SD, 2-3 dayCardiomyocytesCollagenase Type 2: 0.05%DMEMFang, R, Qiao, S., Liu, Y., Meng, Q., Chen, X., Song, B., Hou, X. and Tian, W.: Sustained Co-Delivery of BIO and IGF-1 by a Novel Hybrid Hydrogel System to Stimulate Endogenous Cardiac Repair in Myocardial Infarcted Rat Hearts., Int J Nanomedicine 10, 4691-703, 2015
大鼠大鼠, SDValve Interstitial cellsCollagenase Type 2: 600 u/mlDMEMHorne, T., VandeKopple, M., Sauls, K., Koenig, S., Anstine, L., Garg, V., Norris, R. and Lincoln, J.: Dynamic Heterogeneity of the Heart Valve Interstitial Cell Population in Mitral Valve Health and Disease, J Cardiovasc Dev Dis 2, 214, 2015
大鼠Wistar, 200 gVentricular myocytesCollagenase Type 2: 95 u/mlHEPESMiller, L., Greensmith, D., Sankaranarayanan, R., O'Neill, S. and Eisner D.:The Effect of 2,5-di-(tert-butyl)-1,4-benzohydroquinone (TBQ) on Intracellular Ca2+ Handling in Rat Ventricular Myocytes., Cell Calcium 58, 208-14, 2015
Protease: 0.1 u/ml
大鼠大鼠, SD, 3-4 monthCardiomyocytesCollagenase Type 2: 0.1%DMEMLiu, L., Wang, Y., Cao, Z., Wang, M., Liu, X., Gao, T., Hu, Q., Yuan, W. and Lin, L.: Up-regulated TLR4 in Cardiomyocytes Exacerbates Heart Failure after Long-Term Myocardial Infarction., J Cell Mol Med 19, 2728, 2015
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsHBSSSong, D., Lee, K., Ryu, J., Jeon, H. and Kim, D.: The Molecular Interaction of Heart LIM Protein (HLP) with RyR2 and Caveolin-3 is Essential for Ca(2+)-Induced Ca(2+) Release in the Heart., Biochem Biophys Res Commun 463, 975-81, 2015
大鼠大鼠, neonatalCardiomyocytesTrypsin: 0.06%HBSSZhang, B., Montgomery, M., Davenport-Huyer, L., Korolj, A. and Radisic, M.: Platform Technology for Scalable Assembly of Instantaneously Functional Mosaic Tissues., Sci. Adv. 1, e1500423, 2015
Collagenase Type 2: 220 u/ml
大鼠大鼠CardiomyocytesCollagenase Type 2: 150 u/mlMEMHennessey, J., Wei, E. and Pitt, G.: Fibroblast Growth Factor Homologous Factors Modulate Cardiac Calcium Channels., Circ Res 113, 381-8, 2013
大鼠大鼠, Wistar, 175-200gCardiomyocytesCollagenase Type 2: 150 u/mlsee referenceEckerle, L., Felix, S. and Herda, L.: Measurement of Antibody Effects on Cellular Function of Isolated Cardiomyocytes., J Vis Exp 73, e4237, 2013
大鼠大鼠, SD, 1-2 dayCardiomyocytesTrypsin: 0.06%DMEMZhang, B., Green, J., Murthy, S. and Radisic, M.: Label-Free Enrichment of Functional Cardiomyocytes using Microfluidic Deterministic Lateral Flow Displacement., PLoS ONE 7, e37619, 2012
Collagenase Type 2: 220 u/ml
大鼠大鼠, adultVentricular myocytesCollagenase Type 2: 0.2%Medium 199Karakikes, I., Hadri, L., Rapti, K., Ladage, D., Ishikawa, K., Tilemann, L., Yi, G., Morel, C., Gwathmey, J., Zsebo, K., Weber, T., Kawase, Y. and Hajjar, R.: Concomitant Intravenous Nitroglycerin with Intracoronary Delivery of AAV1.SERCA2a Enhances Gene Transfer in Porcine Hearts., Mol Therapy 20, 565, 2012
大鼠大鼠, 3 dayCardiomyocytesCollagenase Type 2: 300 u/mlPBSYe, K., Sullivan, K. and Black, L.: Encapsulation of Cardiomyocytes in a Fibrin Hydrogel for Cardiac Tissue Engineering., J Vis Exp 55, 3251, 2011
大鼠大鼠, SD, maleCardiomyocytesCollagenase Type 2: 0.056%KrebsMellor, K., Bell, J., Wendt, I., Davidoff, A., Ritchie, R. and Delbridge, L.:Fructose Modulates Cardiomyocyte Excitation-Contraction Coupling and Ca Handling In Vitro., PLoS ONE 6, e25204, 2011
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Dittami, G., Rajguru, S., Lasher, R., Hitchcock, R. and Rabbitt, R.:Intracellular Calcium Transients Evoked by Pulsed Infrared Radiation in Neonatal Cardiomyocytes., J Physiol 589, 1295-306, 2011
大鼠大鼠, neonatalCardiomyocytesTrypsin: 0.07%DMEM/M199Guan, X., Delo, D., Atala, A. and Soker, S.: In Vitro Cardiomyogenic Potential of Human Amniotic Fluid Stem Cells., J Tissue Eng Regen Med 5, 220, 2011
Collagenase Type 2: 0.1%
大鼠大鼠, adultCardiomyocytesCollagenase Type 2: 0.025%see referenceLouch, W., Sheehan, K. and Wolska, B.: Methods in Cardiomyocyte Isolation, Culture, and Gene Transfer., J Mol Cell Cardiol 51, 288-98, 2011
大鼠大鼠, Wistar, maleCardiomyocytesNCIS kit: per instructionsL-15Brinckmann, M., Kaschina, E., Altarche-Xifro, W., Curato, C., Timm, M., Grzesiak, A., Dong, J., Kappert, K., Kintscher, U., Unger, T. and Li, J.:Estrogen Receptor Alpha Supports Cardiomyocytes Indirectly Through Post-Infarct Cardiac c-kit+ Cells., J Mol Cell Cardiol 47, 66, 2009
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Smith, M., Huang, Y. and Deshmukh, M.: Skeletal Muscle Differentiation Evokes Endogenous XIAP to Restrict the Apoptotic Pathway., PLoS ONE 4, e5097, 2009
大鼠大鼠, adultMyocytesCollagenase Type 2: 0.05%Media 199Xu, X. and Colecraft, H.: Primary Culture of Adult Rat Heart Myocytes., J Vis Exp 28, 1308, 2009
Protease XIV: 0.02%
大鼠大鼠, SD, 200-250 gCardiomyocytesCollagenase Type 1: 0.05%Krebs-HenseleitFarkas, A., Acsai, K., Nagy, N., Toth, A., Fulop, F., Seprenyi, G., Birinyi, P., Nanasi, P., Forster, T., Csanady, M., Papp, J., Varrro, A. and Farkas, A.:Na(+)/Ca(2+) Exchanger Inhibition Exerts a Positive Inotropic Effect in the Rat Heart, but Fails to Influence the Contractility of the Rabbit Heart., Br J Pharmacol 154, 93, 2008
Hyaluronidase: see reference
Protease: see reference
大鼠大鼠, SD, 1-2 dayCardiomyocytesNCIS kit: per instructionsL-15Kim, M., Oh, J., Sakata, S., Liang, I., Park, W., Hajjar, R. and Lebeche, D.:Role of Resistin in Cardiac Contractility and Hypertrophy., J Mol Cell Cardiol 45, 270, 2008
大鼠大鼠, SD, 2 dayCardiomyocytesCollagenase Type 2: 0.1%DMEM/F-12Jang, J., Ku, S., Kim, J., Choi, K., Kim, Y., Kim, H., Oh, S., Lee, E., Cho, H., Song, Y., Lee, S., Lee, S., Suh, C., Kim, S., Moon, S. and Choi, Y.: Notch Inhibition Promotes Human Embryonic Stem Cell-Derived Cardiac Mesoderm Differentiation., Stem Cells 26, 2782, 2008
大鼠大鼠, SD, male, 225-250gCardiomyocytesCollagenase Type 2: 0.1%J-MEMKubli, D., Quinsay, M., Huang, C., Lee, Y., Gustafsson, A.: Bnip3 Functions as a Mitochondrial Sensor of Oxidative Stress During Myocardial Ischemia and Reperfusion., Am J Physiol Heart Circ Physiol 295, H2025, 2008
大鼠大鼠, 2-3 dayCardiomyocytesCollagenase Type 2: 0.05%DMEMLaFramboise, W., Scalise, D., Stoodley, P., Graner, S., Guthrie, R., Magovern, J and Becich, M.: Cardiac Fibroblasts Influence Cardiomyocyte Phenotype in Vitro., Am J Physiol Cell Physiol 292, C1799-808, 2007
Pancreatin: 0.1%
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Robinet, A., Millart, H., Oszust, F., Hornebeck, W. and Bellon, G.: Binding of Elastin Peptides to S-Gal Protects the Heart Against Ischemia/Reperfusion Injury by Triggering the RISK Pathway., FASEB J 21, 1968, 2007
大鼠大鼠, SD, 300-350 gCardiomyocytesCollagenase Type 2: 0.05%DMEMNiederbichler Andreas D, Hoesel Laszlo M, Westfall Margaret V, Gao Hongwei, Ipaktchi Kyros R, Sun Lei, Zetoune Firas S, Su Grace L, Arbabi Saman, Sarma J Vidya, Wang StewartC, Hemmila MarkR, Ward PeterA:An essential role for complement C in the pathogenesis of septic cardiac dysfunction, J Exp Med 203, 53-61, 2006
Hyaluronidase: 0.02%
大鼠大鼠, SD, maleVentricular myocytesCollagenase: 0.13%DMEMStagg Mark A, Coppen Steven R, Suzuki Ken, Varela-Carver Anabel, Lee Joon, Brand Nigel J, Fukushima Satsuki, Yacoub Magdi H, Terracciano Cesare M: Evaluation of frequency, type, and function of gap junctions between skeletal myoblasts overexpressing connexin43 and cardiomyocytes: relevance to cell transplantation, FASEB J 20, 744-6, 2006
Hyaluronidase: 0.06%
大鼠大鼠, SD, 200-250 gVentricular myocytes and mesenteric arterial SMCCollagenase Type 2: 0.06%Krebs-RingerLu Tong, Ye Dan, Wang Xiaoli, Seubert John M, Graves Joan P, Bradbury J Alyce, Zeldin Darryl C, Lee Hon-Chi: Cardiac and vascular KATP channels in rats are activated by endogenous epoxyeicosatrienoic acids through different mechanisms, J Physiol 575, 627-44, 2006
Papain: 0.175%
大鼠大鼠, Wistar, neonatalCardiomyocytesNCIS kit: per instructionsL-15Butler, T., Au, C., Yang, B., Egan, J., Tan, Y., Hardeman, E., North, K., Verkman, A. and Winlaw, D.: Cardiac Aquaporin Expression in Humans, Rats, and Mice., Am J Physiol Heart Circ Physiol 291, H705, 2006
大鼠大鼠, 1-2 dayCardiomyocytesNCIS kit: per instructionsDMEM/F-12Pedram, A., Razandi, M., Aitkenhead, M., and Levin, E: Estrogen Inhibits Cardiomyocyte Hypertrophy in vitro. Antagonism of Calcineurin-related Hypertrophy through Induction of MCIP1, J Biol Chem 280, 26339, 2005
大鼠大鼠, SD, male, 250-300 gCardiomyocytesCollagenase Type 2: 250 u/mlCardioplegic solutionHead, B., Pa, H., Roth, D., Lai, N., Niesman, I., Farquhar, M. and Insel, P.: G-Protein-Coupled Receptor Signaling Components Localize in Both Sarcolemmal and Intracellular Caveolin-3-Associated Microdomains in Adult Cardiac Myocytes., J Biol Chem 280, 31036-44, 2005
大鼠大鼠, Wistar, 1-3 dayCardiomyocytesNCIS kit: per instructionsL-15Castillo, A., Ruzmetov, N., Harvey, K., Stillwell, W., Zaloga, G. and Siddiqui, R.: Docosahexaenoic Acid Inhibits Protein Kinase C Translocation/Activation and Cardiac Hypertrophy in Rat Cardiomyocytes., J Mol Genet Med 1, 18, 2005
大鼠大鼠, SD, 2-4 dayCardiomyocytes, fibroblastsTrypsin: 0.1%Medium 199Entcheva, E., Bien, H, Yin L., Chung, C., Farrell, M., and Kostov, Y.:Functional Cardiac Cell Constructs on Cellulose-based Scaffolding,Biomaterials 25(26), 5753, 2004
Collagenase: 0.1%
大鼠大鼠, SD, 1-2 dayCardiomyocytesNCIS kit: per instructionsL-15Muller-Bore, B., Cascio, W., Anderson, P., Snowwaert, J., Frye, J., Desai, N., Esch, G., Brackham, J., Bagnell, C., Coleman, W., Grisham, J., and Malouf, N.: Adult-derived Liver Stem Cells Acquire a Cardiomyocyte Structural and Functional Phenotype ex vivo, Am J Pathol 165, 135, 2004
大鼠大鼠,1-2 dayCardiomyocytesNCIS kit: per instructionsDMEMNatarajan AR, Rong Q, Katchman AN, and Ebert SN.: Intrinsic cardiac catecholamines help maintain beating activity in neonatal rat cardiomyocyte cultures, Pediatr Res 56, 411, 2004
大鼠大鼠, 1 dayCardiomyocytesNCIS kit: per instructionsF-10Chen, Z., Ge, Y., and Kang J.: Down-regulation of the M6P/IGF-II Receptor Increases Cell Proliferation and Reduces Apoptosis in Neonatal Rat Cardiac Myocytes, BMC Cell Biol 5, 15, 2004
大鼠大鼠, neonatalCardiac myocytesCollagenase Type 2: 0.5%HBSSChen Hsiao-Huei, Mullett Steven J, Stewart Alexandre F R: Vgl-4, a novel member of the vestigial-like family of transcription cofactors, regulates alpha1-adrenergic activation of gene expression in cardiac myocytes, J Biol Chem 279, 30800-6, 2004
大鼠大鼠, male, 250-300 gCardiomyocytesCollagenase Type 1: 0.03%HBSSHunton DaciaL, Zou LuYun, Pang Yi, Marchase Richard B: Adult rat cardiomyocytes exhibit capacitative calcium entry, Am J Physiol Heart Circ Physiol 286, H1124-32, 2004
Protease: 0.01%
大鼠大鼠, neonatalCardiomyocytesCollagenase Type 2: 0.1%DMEMTamamori-Adachi, M., Hayashida, K., Nobori, K., Omizu, C., Yamada, K., Sakamoto, N., Kamura, T., Fukuda, K., Ogawa, S., Nakayama, K. and Kitajima, S.: Down-Regulation of p27 Promotes Cell Proliferation of Rat Neonatal Cardiomyocytes Induced by Nuclear Expression of Cyclin D1 and CDK4, J Biol Chem 2279, 50429, 2004
大鼠大鼠, fetal or 3-dayCardiomyocytesCollagenase Type 2: 0.1%HEPESTamamori-Adachi Mimi, Ito Hiroshi, Sumrejkanchanakij Piyamas, Adachi Susumu, Hiroe Michiaki, Shimizu Masato, Kawauchi Junya, Sunamori Makoto, Marumo Fumiaki, Kitajima Shigetaka, Ikeda Masa-Aki: Critical role of cyclin D1 nuclear import in cardiomyocyte proliferation, Circ Res 92, e12-9, 2003
大鼠大鼠, adult, 220-280gCardiomyocytesCollagenase Type 2: 178 u/mlKrebs-Henseleit, CFGordon Jennifer M, Dusting Gregory J, Woodman Owen L, Ritchie Rebecca H: Cardioprotective action of CRF peptide urocortin against simulated ischemia in adult rat cardiomyocytes, Am J Physiol Heart Circ Physiol 284, H330-6, 2003
Hyaluronidase: 0.01%
大鼠大鼠, 50 gVentricular myocytesCollagenase: 223 u/mlMedium 199Aberle II NS, and Ren J.: Experimental Assessment of the Role of Acetaldehyde in Alcoholic Cardiomyopathy, Biol Proced Online 5, 1, 2003
Hyaluronidase: 0.01%
Trypsin: 0.002%
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Li TS, Ito H, Kajiwara K, and Hamano K.: Long-term survival of xenografted neonatal cardiomyocytes by adenovirus-mediated CTLA4-Ig expression and CD40 blockade, Circulation 108, 1760, 2003
大鼠大鼠, SD, 200-300 gCardiomyocytesCollagenase Type 2: 0.05-0.1%Joklik's MEMSwift Luther, McHowat Jane, Sarvazyan Narine: Inhibition of membrane-associated calcium-independent phospholipase A2 as a potential culprit of anthracycline cardiotoxicity, Cancer Res 63, 5992-8, 2003
大鼠大鼠, SD, 175-200gCardiomyocytesCollagenase Type 2: 0.05%Krebs-RingerTardif Annie, Julien Nathalie, Chiasson Jean-Louis, Coderre Lise:Stimulation of glucose uptake by chronic vanadate pretreatment in cardiomyocytes requires PI 3-kinase and p38 MAPK activation, Am J Physiol/Endo 284, E1055-64, 2003
Deoxyribonuclease I: 0.02%
大鼠大鼠, SD, pregnant femalePeritoneal mastHyaluronidase: 100 u/mlDMEM (see reference)DeAlmeida, A., Mustin, D., Forman, M., Brower, G., Janicki, J., and Carver, W.: Effects of mast cells on the behavior of isolated heart fibroblasts: modulation of collagen remodeling and gene expression, J Cell Physiol 191, 51, 2002
大鼠大鼠, neonatal, adult, 17 dayVentricular cardiomyocytesCollagenase Type 2: 0.7-1%DMEMLam ML, Bartoli M, and Claycomb WC.: The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression, Mol Cell Biochem 229, 51, 2002
大鼠大鼠, 1 dayCardiomyocytesNCIS kit: per instructionsL-15Yang Y, Liao H, Ke Q, Cai J, Xiao YF, and Morgan JP: Enhancement of nitric oxide production by methylecgonidine in cultured neonatal rat cardiomyocytes, Br J Pharmacol 135, 188, 2002
大鼠大鼠, female, 250-300 gVentricular myocytesCollagenase Type 2: 0.1%Ringer solutionLiu Qinghang, Hofmann Polly A: Antiadrenergic effects of adenosine A(1) receptor-mediated protein phosphatase 2a activation in the heart, Am J Physiol Heart Circ Physiol 283, H1314-21, 2002
大鼠大鼠, Wistar, 1 dayCardiomyocytesCollagenase Type 2: 80u/mlM199Shimizu Tatsuya, Yamato Masayuki, Isoi Yuki, Akutsu Takumitsu, Setomaru Takeshi, Abe Kazuhiko, Kikuchi Akihiko, Umezu Mitsuo, Okano Teruo: Fabrication of pulsatile cardiac tissue grafts using a novel 3-dimensional cell sheet manipulation technique and temperature-responsive cell culture surfaces, Circ Res 90, e40, 2002
大鼠大鼠, male, 6-12 months oldCardiac myocytesProtease: 0.55 u/mlTyrode's solutionKitta, K., Day, R., Ikeda, T., and Suzuki, Y.: Hepatocyte Growth Factor Protects Cardiac Myocytes Against Oxidative Stress-Induced Apoptosis, Free Radic Biol Med 31 (7), 902, 2001
大鼠大鼠, Harlan SD, pupCardiomyocytesTrypsin: DMEM/Ham's F-12Schwartzbauer, G., and Robbins, J.: The Tumor Suppressor Gene PTEN Can Regulate Cardiac Hypertrophy and Survival, J Biol Chem 276, 35786, 2001
大鼠大鼠, SD, newbornCardiac myocytesNCIS kit: per instructionsL-15Rahman A, Alam M, Rao S, Cai L, Clark LT, Shafiq S, and Siddiqui MA:Differential effects of doxorubicin on atrial natriuretic peptide expression in vivo and in vitro, Biol Res 34(3-4), 195, 2001
大鼠大鼠, adultVentricular myocytesCollagenase: 223 u/mlTyrode solutionRen J, and Wold LE.: Measurement of Cardiac Mechanical Function in Isolated Ventricular Myocytes from Rats and Mice by Computerized Video-Based Imaging, Biol Proced Online 3, 43, 2001
Hyaluronidase: 0.01%
Trypsin: 0.002%
大鼠大鼠, SD, maleCardiomyocytesCollagenase Type 2: 140 u/mlKrebs-HenseleitDai L, Brookes PS, Darley-Usmar VM, and Anderson PG.: Bioenergetics in cardiac hypertrophy: mitochondrial respiration as a pathological target of NO, Am J Physiol/Heart 281, H2261-H2269, 2001
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsDMEM/M199Guo K, Searfoss G, Krolikowski D, Pagnoni M, Franks C, Clark K, Yu KT, Jaye M, Ivashchenko Y: Hypoxia induces the expression of the pro-apoptotic gene BNIP3, Cell Death Differ 8, 367-76, 2001
大鼠大鼠, Wistar, femaleVentricular myocytesCollagenase Type 2: 0.1%Ringer solutionPyle, W, Lester, J, and Hofmann, P.: Effects of Kappa-opioid receptor activation of myocardium, Am J Physiol Heart Circ Physiol 281, H669, 2001
大鼠大鼠, SD, 1-2 dayCardiomyocytesTrypsin: 0.01%HBSSArutunyan A, Webster D, Swift L, Sarvazyan N: Localized injury in cardiomyocyte network: a new experimental model of ischemia-reperfusion arrhythmias, Am J Physiol Heart Circ Physiol 280, H1905-15, 2001
Collagenase Type 2: 0.08%
大鼠大鼠, newbornAtrialTrypsin: 0.06%DMEMKim, D., and Pleumsamran, A.: Cytoplasmic Unsaturated Free Fatty Acids Inhibit ATP-Dependent Gating of the G Protein-gated K+ Channel, J Gen Physiol 115, 287, 2000
大鼠大鼠, 1-2 dayCardiomyocytesCollagenase Type 1: 84 u/mlM199 mediumBueno OF, De Windt LJ, Tymitz KM, Witt SA, Kimball TR, Klevitsky R, Hewett TE, Jones SP, Lefer DJ, Peng CF, Kitsis RN, and Molkentin JD.: The MEK1-ERK1/2 signaling pathway promotes compensated cardiac hypertrophy in transgenic mice, EMBO J 19(23), 6341, 2000
Pancreatin: 0.05%
大鼠大鼠, Winstar, 1-2 dayVentricular myocytesCollagenase Type 2: 0.08%DMEMMaki T, Horio T, Yoshihara F, Suga S, Takeo S, Matsuo H, and Kangawa K: Effect of neutral endopeptidase inhibitor on endogenous atrial natriuretic peptide as a paracrine factor in cultured cardiac fibroblasts, Br J Pharmacol 131(6), 1204, 2000
大鼠大鼠, SD, adult femaleHeart microvascular cellsCLSPA: 250u/mlL-15Puri S, Bansal DD, Uskokovic MR, and MacGregor RR.: Induction of tissue plasminogen activator secretion from rat heart microvascular cells by fM 1,25(OH)(2)D(3), Am J Physiol/Endo 278(2), E293, 2000
Papain: 5 u/ml
Elastase: 0.8 u/ml
大鼠大鼠, SD, male, 250-300gCardiomyocytesCollagenase Type 2: 0.06%M199Sun L, Chang J, Kirchhoff SR, and Knowlton AA.: Activation of HSF and selective increase in heat-shock proteins by acute dexamethasone treatment, Am J Physiol Heart Circ Physiol 278(4), H1091, 2000
大鼠大鼠, wistar, 2 dayCardiomyocytesNCIS kit: per instructionsDMEM-F12Cowan DB, Poutias DN, Del Nido PJ, and McGowan FX Jr: CD14-independent activation of cardiomyocyte signal transduction by bacterial endotoxin, Am J Physiol Heart Circ Physiol 279, H619, 2000
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Webster DR, and Patrick DL.: Beating rate of isolated neonatal cardiomyocytes is regulated by the stable microtubule subset, Am J Physiol Heart Circ Physiol 278, H1653, 2000
大鼠大鼠, Lewis, adult male, 300-350-gAorta, smooth muscleTrypsin: 0.025%Medium 199 with 20% FBS mediumKim, B., Nikolovski, J., Bonadio, J., Smiley, E., and Mooney, D.: Engineered Smooth Muscle Tissues: Regulating Cell Phenotype with the Scaffold, Exp Cell Res 251, 318, 1999
大鼠大鼠, 10-15 newborn, 1-2 days oldAtrialTrypsin: 0.03%HBSS or EBSSNemec, J., Wickman, K., and Clapham, D.: GBy Binding Increases the Open Time of IKACh: Kinetic Evidence for Multiple GBy Binding Sites, Biophys J 76, 246, 1999
大鼠大鼠, 1-3 dayNeonatal rat cardiomyocytesCollagenase Type 2: 80u/mlDMEM/F12Adderley SR, and Fitzgerald DJ.: Oxidative damage of cardiomyocytes is limited by extracellular regulated kinases 1/2-mediated induction of cyclooxygenase-2, J Biol Chem 274, 5038, 1999
Pancreatin: 0.06%
大鼠大鼠, 1-2 dayCardiomyocytesPancreatin: 0.06%DMEMReinecke H, Zhang M, Bartosek T, and Murry CE.: Survival, integration, and differentiation of cardiomyocyte grafts: a study in normal and injured rat hearts, Circulation 100(2), 193, 1999
Collagenase Type 2: 95 u/ml
大鼠大鼠, adult and neonatalCardioomyocytesCollagenase Type 4: 0.12%-adultDMEM/medium 199Richards SM, Jaconi ME, Vassort G, and Puceat M.: A spliced variant of AE1 gene encodes a truncated form of Band 3 in heart: the predominant anion exchanger in ventricular myocytes, J Cell Sci 112, 1519, 1999
Collagenase Type 2: 0.05%-neonatal
大鼠大鼠, Wistar, maleCardiomyocytesCollagenase Type 2: 228 u/mlJoklik MEMSambandam N, Abrahani MA, St Pierre E, Al-Atar O, Cam MC, and Rodrigues B.: Localization of lipoprotein lipase in the diabetic heart: regulation by acute changes in insulin, Arterioscler Thromb Vasc Biol 19(6), 1526, 1999
大鼠大鼠, 1-2 dayCardiomyocytesNCIS kit: per instructionsRPMIMatsui, T., Li, Ling, Del Monte, F., Fukui, Y., Franke, T., Hajjar, R., Rosenzweig, A.: Adenoviral gene transfer of activated phosphatidylinositol 3'-kinase and Akt inhibits apoptosis of hypoxic cardiomyocytes in vitro,Circulation 100, 2373-9, 1999
大鼠大鼠Thoracic aorta, smooth muscleElastase: 0.02%(see reference)Zilberman, A., Dave, V., Miano, J., Olson, E., and Periasamy, M.:Evolutionary Conserved Promoter Region Containing CArg-Like Elements is Crucial for Smooth Muscle Myosin Heavy Chain Gene Expression, Circ Res 82, 566, 1998
大鼠大鼠, SD, 1 dayCardiomyocytesNCIS kit: per instructionsDMEM/F-12Wagner DR, Combes A, McTiernan C, Sanders VJ, Lemster B, Feldman AM: Adenosine inhibits lipopolysaccharide-induced cardiac expression of tumor necrosis factor-alpha, Circ Res 82, 47-56, 1998
大鼠大鼠, SD, 300-350 gCardiomyocytesCollagenase Type 2: 0.04%see referenceYu L, Netticadan T, Xu YJ, Panagia V, Dhalla NS: Mechanisms of lysophosphatidylcholine-induced increase in intracellular calcium in rat cardiomyocytes, J Pharmacol Exp Ther 286, 1-8, 1998
大鼠大鼠, SD, female, 200gVentricular cardiac myocytesCollagenase Type 2: 0.5%Krebs-Henseleit bufferWestfall M V, Rust E M, Metzger J M: Slow skeletal troponin I gene transfer, expression, and myofilament incorporation enhances adult cardiac myocyte contractile function, Proc Natl Acad Sci U S A 94, 5444-9, 1997
Hyaluronidase: 0.2%
大鼠大鼠, Wistar, 1 dayCardiomyoctesCollagenase Type 2: 80 u/mlHBSSKinugawa, K., Kohmoto, O., Yao, A., Serizawa, T., and Takahashki, T.:Cardiac inducible nitric oxide synthase negatively modulates myocardial function in cultured rat myocytes, Am J Physiol 272, H35-47, 1997
大鼠大鼠, neonatal, 5-dayCardiomyocytesTrypsin: 0.05%DMEMVanwinkle, W., Snuggs, M., and Buja, L.: Cardiogel: A Biosynthetic Extracellular Matrix for Cardiomyocyte Culture, In Vitro Cell Dev Biol Anim 32, 478, 1996
大鼠大鼠, SD, 250-300 gCardiomyocytesCollagenase Type 2: 0.05%Joklik'sSharma VK, Colecraft HM, Wang DX, Levey AI, Grigorenko EV, Yeh HH, and Sheu SS: Molecular and functional identification of m1 muscarinic acetylcholine receptors in rat ventricular myocytes, Circ Res 79(1), 86, 1996
大鼠大鼠, SD, fetal and neonatalCardiomyocytesCollagenase: 0.1%PBSLi RK, Mickle DA, Weisel RD, Zhang J, Mohabeer MK: In vivo survival and function of transplanted rat cardiomyocytes, Circ Res 78, 283-8, 1996
Trypsin: 0.2%
大鼠SD, SHR or WKYCardiomyocytesNCIS kit: per instructionsHBSSMacGregor, R., Klein, R., and Bansal, D.: Secretion of Plasminogen Activator Activity from Neonatal Rat Heart Cells is Regulated by Hormones and Growth Factors, Ann N Y Acad Sci 752, 331, 1995
大鼠大鼠, SD, male, 200-250 gMyocytesCollagenase: 0.07%Joklik's MEMLaughlin, M., Schaeffer, M., and Sturek, M.: Effect of Exercise Training on Intracellular Free Ca2+ Transients in Ventricular Myocytes of Rats, Am J Physiol 73, 1141, 1992
Ventricles
大鼠大鼠, SD, female, 60-90 daysMyocardialCollagenase Type 2: 0.1%Joklik's MEM, Kreb's-Henseleit buffer, CFWelder, A.A., Grant, R., Bradlaw, J., and Acosta, D.: A Primary Culture System of Adult Rat Heart Cells for the Study of Toxicologic Agents, In Vitro Cell Dev Biol 27, 921, 1991
大鼠大鼠, SD, adult, female, 180-220 gCardiomyocytesCollagenase: 0.11%(see reference)Fischer, Y., Rose, H., and Kammermeier. H.: Highly Insulin-Responsive Isolated Rat Heart Muscle Cells Yielded By a Modified Isolation Method, Life Sci 49, 1679, 1991
大鼠大鼠, neonatal, 1-4 daysCardiomyocytesCollagenase Type 2: 0.12%Krebs-Henseleit Buffer, CFMarino, T.A., Walter, R.,Cobb, E., Palasiuk, M., Parsons, T., and Mercer, W.E.: Effects of Norepinephrine on Neonatal Rat Cardiocyte Growth and Differentiation, In Vitro Cell Dev Biol 26, 229, 1990
大鼠大鼠 HeartTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and B綿羊 Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
(also b綿羊)Adrenal chromaffin 
 Paraneurons
大鼠大鼠, adult, femaleMyocytesCollagenase Type 2: 0.05%Joklik's MEMNag, A.C., Lee, M., and Kosiur, J.R.: Adult Cardiac Muscle Cells in Long-Term Serum-Free Culture:Myofibrillar Organization and Expression of Myosin Heavy Chain Isoforms, In Vitro Cell Dev Biol 26, 464, 1990
大鼠大鼠s, AW, late-gestation pregnant femalesVentricularCollagenase Type 2: 0.05 - 0.08%See referenceEngelmann, G., McTiernan, C., Gerrity, R., and Samarel, A.: Technique 2 (6), 279, 1990
Cardiomyocytes
大鼠大鼠,SD, albino, 10-14 weeksMyocytesCollagenase Type 1: 100 u/ml and 150 u/mlKrebs Ringer bicarbonate bufferBerg, I., Guse, A.H., and Gercken, G.: Carbamoylcholine-Induced Accumulation of Inositol Mono-,Bis-,Tris-, And Tetrakisphosphates In Isolated Cardiac Myocytes From Adult Rats, Biochim Biophys Acta 1010, 100, 1989
大鼠大鼠, SD, adult, female, 175 - 225 gMyocytesCollagenase: 0.1%Joklik's mediumBugaisky, L.B. and Zak, R.: Differentiation of Adult Rat Cardiac Myocytes in Cell Culture, Circ Res 64, 493, 1989
大鼠大鼠, SD, male, 250 - 350 gMyocytes, ventricularCollagenase Type 1: 90 and 100 u/mlJoklik's solution with and without calciumDe Young, M., Giannattasio, B., and Scarpa, A.: Isolation of Calcium-Tolerant Atrial And Ventricular Myocytes From Adult Rat Heart,Methods Enzymology Vol. 173,, , 662, 1989
大鼠大鼠, SD, 2-4 day oldCardiomyocytesTrypsin: 0.1%HBSSToraason, M., Luken, M., Breitenstein, M., Krueger, J., and Biagini, R.:Comparative Toxicity of Allylamine and Acrolein in Cultured Myocytes and Fibroblasts from Neonatal Rat Heart, Toxicology 56, 107, 1989
Fibroblasts
大鼠大鼠, SD, maleAortic smooth muscleTrypsin: 0.05%DMEMCornwell, T., and Lincoln, T.: Regulation of Intracellular Ca2+ Levels in Cultured Vascular Smooth Muscle Cells, J Biol Chem 264 (2), 1146, 1989
大鼠大鼠, adult, 150-200 gMyocytes, atriaCollagenase Type 1: 100 u/mlKreb's Ringer bicarbonate - HEPES bufferMcMahon, K.K.: A Study of Adult Rat Atrial Myocyte Attachment to Extricellular Matrix Components and Long Term Culture,Biology of Isolated Adult Cardiac Myocytes,(Clark, Decker, Borg, Eds), Elsevier Science Publishing Co. Inc., 318, 1988
大鼠SD大鼠MyocytesCollagenase Type 2: 0.1%Kreb's Ringer w/ calciumBuxton, I.L.O. and Doggwiler, K.O.: Alpha1-Adrenergic Receptor Signal Transduction in the Adult Rat Cardiac Myocyte,Biology of Isolated Adult Cardiac Myocytes,(Clark, Decker, Borg, eds), , 298, 1988
大鼠SD大鼠, 雄性, 250-350 克Myocytes, heartHyaluronidase: 0.03%Bicarbonate-buffered mediumKim, D. and Smith, T.W.: Temporal Variation in Contractile State And [Ca++] in Isolated Adult Rat And Guinea Pig Cardiac Myocytes,Biology of Isolated Adult Cardiac Myocytes,(Clark, Decker, Borg, eds), Elsevier Science Publishing Co. Inc., 370, 1988
大鼠Wistar大鼠,  2-4 天neonatesMyoblast, cardiacTrypsin: 0.1%Standardized MediumGrynberg, A., Degois, M., Guenot, L., and Athias, P.: Primary Rat Cardiac Cell Culture: Diet of the Mother Rats as a Determinant Parameter of Cardiomyoblast Production from Neonates, Biol Cell 57, 89, 1986
(see reference)
大鼠大鼠, SD, 懷孕的MyocytesTrypsin: 2.4 u/mlDMEMFreerksen, D., Schroedl, N., and Hartzell, C.: Control of Enzyme Activity Levels by Serum and Hydrocortisone in Neonatal Rat Heart Cells Cultured in Serum-Free Medium, J Cell Physiol 120, 126, 1984
大鼠大鼠, SD, 雌性, 12 周MyocytesCollagenase Type 1: 0.06%Krebs Ringer bicarbonate bufferPiper, H.M., Probst, I., Schwartz, P., Hutter, F.J., and Spieckermann, P.G.:Culturing of Calcium Stable Adult Cardiac Myocytes, J Mol Cell Cardiol 14, 397, 1982
大鼠大鼠, 雄性/雌性, 200-250 克MuscleHyaluronidase: 0.1%Krebs Ringer phosphate buffer, CMFNag, A. and Cheng, M.: Adult Mammalian Cardiac Muscle Cells in Culture,Tissue Cell 13, 515, 1981
大鼠大鼠, 雌性, 成年, 200 - 300 克MyocytesCollagenase Type 2: 0.1%Joklik's MEMClaycomb, W.C., and Palazzo, M.C.: Culture of The Terminally Differentiated Adult Cardiac Muscle Cell: A Light and Scanning Electron Microscope Study,Dev Biol 80, 466, 1980
大鼠SD大鼠, 成年, 雄性, 200-250 克MyocytesHyaluronidase: 0.10%Joklik MEM, CFFrangakis, C., Bahl, J., McDaniel, H., and Bressler, R.: Tolerance to Physiological Calcium by Isolated Mycocytes from the Adult Rat Heart; An Improved Cellular Preparation, Life Sci 27, 815, 1980
大鼠SD大鼠, 雌性, 12 周, 250 克MyocytesCollagenase Type 1: 0.20%Kreb's Ringer bicarbonate bufferPowell, T., Terrar, D.A., and Twist, V.W.: Electrical Properties of Individual Cells Isolated From Adult Rat Ventricular Myocardium, J Physiol 302, 131, 1980
Ventricular myocardium
大鼠大鼠, 10 周MyocytesCollagenase Type 1: 0.1%SalineBishop, S., Oparil, S., Reynolds, R., and Drummond, J.: Regional Myocyte Size in Normotensive and Spontaneously Hypertensive Rats, Hypertension 1 (4), 378, 1979
大鼠大鼠, 雄性, 200-300 克MyocytesHyaluronidase: 82 u/mlKreb's Henseleit bufferFarmer, B., Harris, R., Jolly, W., Hathaway, D., Katzberg, A., Watanabe, A., Whitlow, A., and Besch, H.: Isolation and Characterization of Adult Rat Heart Cells, Arch Biochem Biophys 179, 545, 1977
大鼠大鼠,雌性, 白化變種, 200-250克MyocytesHyaluronidase: 0.1%Kreb's Ringers phosphateNag, A., Fischman, D., Aumont, M., and Zak, R.: Studies of Isolated Adult Rat Heart Cells: The Surface Morphology and the Influence of Extracellular Calcium Ion Concentration on Cellular Viability, Tissue Cell 9 (3), 419, 1977
大鼠大鼠, 成年, 雄性, 150-200克MyocytesCollagenase: 0.1%MEM CFGrosso, D., Frangakis, C., Carlson, E., and Bressler, R.: Isolation and Characterization of Myocytes From the Adult Rat Heart, Prep Biochem 7 (5), 383, 1977
Hyaluronidase: 0.1%
大鼠大鼠, 成年的MyocytesCollagenase: 0.1%Perfusing solutionMoustafa, E., Skomedal, T., Osnes, J., and Oye, I.: Cyclic Amp Formation and Morphology of Myocardial Cells Isolated from Adult Heart: Effect of Ca2+ and Mg2+, Biochim Biophys Acta 421, 411, 1976
大鼠 SD大鼠, 白化變種, 雄性, 180-230 克Heart ventricles, beatingCollagenase: 0.05-0.2%Phosphate bufferGlick, M., Burns, A., and Reddy, W.: Dispersion and Isolation of Beating Cells from Adult Rat Heart, Anal Biochem 61, 32, 1974
大鼠大鼠, 希伯來聯(lián)合國, 3 & 17 個月, 150-210 克MyocytesTrypsin: 0.05%Versene bufferBierman, E.L., Stein, O., and Stein, Y.: Lipoprotein Uptake and Metabolism by Rat Aortic Smooth Muscle Cells in Tissue Culture, Circ Res 35, 136, 1974
大鼠大鼠, 2-5 天MyocytesTrypsin: 0.01%Saline ASpeicher, D., and McCarl, R.: Pancreatic Enzyme Requirements for the Dissociation of Rat Hearts for Culture, In Vitro 10, 30, 1974
大鼠大鼠, SD, male, 150-200克MyocytesHyaluronidase: 0.05%(see reference)Pretlow II, T., Glick, M., and Reddy, W.: Separation of Beating Cardiac Myocytes from Suspensions of Heart Cells, Am J Pathol 67 (2), 215, 1972
大鼠大鼠, albino, male, 250 克MyocytesHyaluronidase: 0.20%Hank's solution, CFBerry, M., Friend, D., and Scheuer, J.: Morphology and Metabolism of Intact Muscle Cells Isolated from Adult Rat Heart, Circ Res 26, 679, 1970
大鼠大鼠, 成年的, 雄性, 250 克MyocytesTrypsin: 0.1%Saline AVahouny, G.V., Wei, R., Starkweather, R., and Davis, C.: Preparation of Beating Heart Cells From Adult Rats, Science 167, 1616, 1970
大鼠Wistar大鼠,  1-4 天大MyocytesTrypsin: 0.125%Gey's BSSMark, G., and Strasser, F.: Pacemaker Activity and Mitosis in Cultures of Newborn Rat Heart Ventricle Cells, Exp Cell Res 44, 217, 1966
貝類動物太平洋牡蠣HaemocytesTrypsin: L15 mediumLe Duff, R., Lipart, C., and Renault, T.: Primary Culture of Pacific Oyster, Crassostrea gigas, heart cells, J Tiss Cul Meth 16, 67, 1994
貝類動物蚌, 3.0 - 4.5 厘米MyocytesCollagenase Type 1: 2%L15 mediumWen, C., Kou, G., and Chen, S.: C*tion of Cells From the Heart of the Hard Clam, Merretrix lusoria (Roding), J Tiss Cul Meth 15, 123, 1993
Fibroblasts

 

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6:我們還是Santa,Advanced Biotechnologies Inc, Worthington Biochemical ,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. Dako,Inc, GePromegaridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,Worthington Biochemical , Polysciences,prospecbi, QA-BIO,quickzyme,FULLERLABS,INC,sterlitech;sysy,TriLink BioTechnologies,Inc;worthington-biochem,zyagen等幾十家國外公司授權代理。
7:我們還是invitrogen,qiagen,Worthington Biochemical am,sigma;Promega,roche,merck, rnd,BD, GE,pierce,BioLegend等*批發(fā),歡迎合作。

 

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