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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
Bovine (also rat)心脏,肾上腺嗜铬,副神经元 / Heart,Adrenal chromaffin,ParaneuronsTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, 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 / 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/狗Canine, adult心房和心室肌细胞 / Atrial 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
犬/狗Canine心肌细胞 / CardiomyocytesCollagenase Type 2: 300 u/ml;
Protease: 0.03%
Tyrode'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
犬/狗Canine心房肌细胞 / 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
犬/狗Canine心肌细胞 / CardiomyocytesCollagenase Type 2: 60 u/ml;
Protease XIV: 0.5 u/ml
See 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
犬/狗Canine心房和心室肌细胞 / 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/狗Canine, adult心肌细胞 / CardiomyocytesCollagenase Type 2: 0.05%;
Protease Type XIV: 0.01%
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
犬/狗Canine, 20-25 kg心肌细胞 / CardiomyocytesCollagenase 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/狗Canine, adult心肌细胞 / CardiomyocytesCollagenase Type 2: 0.05%;
Protease: 0.008%
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
犬/狗Canine心房和心室肌细胞 / 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
犬/狗Canine心房肌细胞 / 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/狗Canine, 20-30 kg心肌细胞 / CardiomyocytesCollagenase Type 2: 110 u/mlTyrode's solutionSun, 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 myocytesCollagenase Type 4: 0.04%;
Collagenase Type 2: 125 u/ml
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
犬/狗Dog, mongrel, adult肌细胞 / MyocytesHyaluronidase: 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
Chicken, embryonic心肌细胞 / CardiomyocytesTrypsin: 0.13%;
Collagenase Type 2: 0.13%;
Deoxyribonuclease I: 0.033%
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
Chicken embryos, 9-11 day incubated心肌细胞 / CardiomyocytesTrypsin: 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
Chick, embryos, 10-11 day心肌细胞 / CardiomyocytesTrypsin: 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
Chick, 10 day心室 / VentricularTrypsin: 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
Chick, embryo, 11 day心肌细胞 / CardiomyocytesTrypsin: 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
Chick, embryonic, 11-12-day-old肌细胞 / MyocytesTrypsin: 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
Chick, 8 day embryo (also rat, neonate and mouse)肌细胞 / 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
Hen, White Leghorn心脏 / HeartTrypsin: 0.025%Medium 199Dehann, R.: Regulation of Spontaneous Activity and Growth of Embryonic Chick Heart Cells in Tissue Culture, Dev Biol 16, 216, 1967
Chick embryos, 5 day心,肝 / Heart,LiverTrypsin: 3.0%Tyrode's solution, CMFSteinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res 30, 257, 1963
Cat, mongrel, adult, 1.8-2.8 Kg肌细胞,心室 / Myocytes,VentricularCollagenase 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
Zebrafish心肌细胞 / Cardiac MyocytesProtease XIV: 0.005%;
Collagenase Type 1: 0.1-0.2%;
Trypsin: 0.08%
HEPESFowler, 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
Fish, Danio rerio心肌细胞 / CardiomyocytesCollagenase Type 2: 0.5%;
Collagenase Type 4: 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
青蛙Frog (Rana esculenta)肌细胞 / 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, male, 350-400g心室肌细胞 / Ventricular myocytesCollagenase Type 2: 0.4%Tyrodes solutionWang, 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 / CardiomyocytesCollagenase 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Guinea-Pig心肌细胞 / 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, 300-350g冠状动脉内皮 / Coronary 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 g内皮 / EndothelialCollagenase 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


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