worthington 心脏组织分离指南

发布者：艾美捷科技发布时间：2022-06-07

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原代细胞分离中，针对上皮细胞组织涉及到心脏，心肌细胞、心房、肾上腺嗜铬，副神经元等等的解离通常会使用酶，如何选择合适的酶来完成对应心脏细胞组织的解离，以及原代细胞采集的常规操作， 艾美捷针对心脏细胞相关的组织分类解离分种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献，详细汇总指南：心脏组织分离指南

本技术指南描述了标准实验室程序；为建立细胞分离方案提供了一种合乎逻辑的实验方法；并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南。

心脏
种属	种属详情	细胞名称	酶组合推荐，含推荐浓度	培养基	参考文献
牛	Bovine (also rat)	心脏，肾上腺嗜铬，副神经元 / Heart，Adrenal chromaffin，Paraneurons	Trypsin: 0.06%	25mM HEPES buffered Locke's solution, CMF	Trifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and Bovine Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
犬/狗	Canine	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 150 u/ml	Tyrode's solution	Zhang,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/狗	Canine, adult	心房和心室肌细胞 / Atrial and ventricular myocytes	Collagenase Type 2: 0.065%	See reference	Bonilla, 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
犬/狗	Canine	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 300 u/ml； Protease: 0.03%	Tyrode's solution	Xi, 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
犬/狗	Canine	心房肌细胞 / Atrial myocytes	Collagenase Type 2: 0.03%	Tyrode's solution	Gan, 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
犬/狗	Canine	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 60 u/ml； Protease XIV: 0.5 u/ml	See reference	Harleton, 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
犬/狗	Canine	心房和心室肌细胞 / Atrial and ventricular myocytes	Collagenase Type 2: 0.05-0.08%	See reference	Zhang, 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/狗	Canine, adult	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 0.05%； Protease Type XIV: 0.01%	HEPES	Calloe, 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
犬/狗	Canine, 20-25 kg	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 0.05%	Tyrode's solution	Burashnikov, 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/狗	Canine, adult	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 0.05%； Protease: 0.008%	M199	Gavi, 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
犬/狗	Canine	心房和心室肌细胞 / Atrial and ventricular myocytes	Collagenase Type 2: 0.065%	Tyrode's solution	Schotten, 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
犬/狗	Canine	心房肌细胞 / Atrial myocytes	Collagenase Type 2: 0.013%	HEPES	Baba, 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/狗	Canine, 20-30 kg	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 110 u/ml	Tyrode's solution	Sun, H., Chartier, D., Leblanc, N. and Nattel, S.: Intracellular Calcium Changes and Tachycardia-Induced Contractile Dysfunction in Canine Atrial Myocytes., Cardiovasc Res 49, 751-61, 2001
犬/狗	Canine	心室和心房肌细胞 / Ventricular and atrial myocytes	Collagenase Type 4: 0.04%； Collagenase Type 2: 125 u/ml	HEPES/Tyrode's	Gintant, 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
犬/狗	Dog, mongrel, adult	肌细胞 / Myocytes	Hyaluronidase: 0.1%	Joklik's MEM	Spanier, A. and Weglicki, W.: Ca2+-Tolerant Adult Canine Myocytes: Preparation and Response to Anoxia/Acidosis, Am J Physiol 243, H448, 1982
鸡	Chicken, embryonic	心肌细胞 / Cardiomyocytes	Trypsin: 0.13%； Collagenase Type 2: 0.13%； Deoxyribonuclease I: 0.033%	HBSS	Blech-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
鸡	Chicken embryos, 9-11 day incubated	心肌细胞 / Cardiomyocytes	Trypsin: 0.25%	PBS	Eschenhagen, 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
鸡	Chick, embryos, 10-11 day	心肌细胞 / Cardiomyocytes	Trypsin: 0.17%	HBSS, CMF	Wang, 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
鸡	Chick, 10 day	心室 / Ventricular	Trypsin: 0.025%	HBSS, CMF	Kim, 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
鸡	Chick, embryo, 11 day	心肌细胞 / Cardiomyocytes	Trypsin: 0.05%	HBSS, CMF	Jacob, 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
鸡	Chick, embryonic, 11-12-day-old	肌细胞 / Myocytes	Trypsin: 0.025%	CMF solution	Murphy, 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
鸡	Chick, 8 day embryo (also rat, neonate and mouse)	肌细胞 / Myocytes	Trypsin NF 1:250: 0.25%	Rinaldini's buffer solution, CF	Gross, 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
鸡	Hen, White Leghorn	心脏 / Heart	Trypsin: 0.025%	Medium 199	Dehann, R.: Regulation of Spontaneous Activity and Growth of Embryonic Chick Heart Cells in Tissue Culture, Dev Biol 16, 216, 1967
鸡	Chick embryos, 5 day	心，肝 / Heart，Liver	Trypsin: 3.0%	Tyrode's solution, CMF	Steinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res 30, 257, 1963
猫	Cat, mongrel, adult, 1.8-2.8 Kg	肌细胞，心室 / Myocytes，Ventricular	Collagenase Type 2: 0.12%	Kreb's Henseleit, CF	Silver, L., Hemwall, E., Marino, T., and Houser, S.: Isolation and Morphology of Calcium-Tolerant Feline Ventricular Myocytes, Am J Physiol 245, H891, 1983
鱼	Zebrafish	心肌细胞 / Cardiac Myocytes	Protease XIV: 0.005%； Collagenase Type 1: 0.1-0.2%； Trypsin: 0.08%	HEPES	Fowler, E., Wang, N., Hezzell, M., Chanoit, G., Hancox, J. and Cannell, M.: Arrhythmogenic Late Ca2+ Sparks in Failing Heart Cells and Their Control by Action Potential Configuration, Proc Natl Acad Sci U S A 117, 2687-2692, 2020
鱼	Fish, Danio rerio	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 0.5%； Collagenase Type 4: 0.5%	MEM	Sander, 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
青蛙	Frog (Rana esculenta)	肌细胞 / Myocytes	Trypsin: 0.04%	CF Ringer	Arrio-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, male, 350-400g	心室肌细胞 / Ventricular myocytes	Collagenase Type 2: 0.4%	Tyrodes solution	Wang, Y., Geng, J., Jiang, M., Li, C., Han, Y and Jiang, J.: The Cardiac Electrophysiology Effects of Higenamine in Guinea Pig Heart, Biomed Pharmacother 109, 2348, 2019
豚鼠	Guinea pig, adult	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 100 u/ml	M-199	Zorn-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	Guinea-Pig	心肌细胞 / Cardiomyocytes	Collagenase Type 2: 100-200 u/ml	See reference	Dhamoon, 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, 300-350g	冠状动脉内皮 / Coronary endothelial	Collagenase Type 2: 0.1%	see reference	Buxton 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 g	内皮 / Endothelial	Collagenase Type 2: 0.15%	Perfusing solution (see reference)	Preisig-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