worthington 肌肉組織分離指南

發(fā)布者：艾美捷科技發(fā)布時間：2022-06-07

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原代細(xì)胞分離中，針對肌肉組織涉及到肌肉衛(wèi)星細(xì)胞，肺動脈內(nèi)皮細(xì)胞和平滑肌細(xì)胞、平滑肌、血管平滑肌等等的解離通常會使用酶，如何選擇合適的酶來完成對應(yīng)肌肉組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對肌肉相關(guān)的組織分類解離分種屬、不同的細(xì)胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻，詳細(xì)匯總指南： worthington肌肉組織分離指南

本技術(shù)指南描述了標(biāo)準(zhǔn)實驗室程序；為建立細(xì)胞分離方案提供了一種合乎邏輯的實驗方法；并列出了許多組織特異性參考資料，以幫助開發(fā)有效的方法。更多其他組織的解離指南，點擊查看：各組織分離指南。

肌肉
種屬	種屬詳情	細(xì)胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻
牛	Bovine	肌肉衛(wèi)星細(xì)胞 / Muscle satellite cells	Collagenase Type 2: 300 u/ml	Krebs-Ringer Bicarbonate	Lee, E., Choi, J., Hyun, J., Cho, K., Hwang, I., Lee, H., Chang, J. and Choi, I.: Steroid Effects on Cell Proliferation, Differentiation and Steroid Receptor Gene Expression in Adult Bovine Satellite Cells., AJAS 20, 501, 2007
牛	Bovine	肺動脈內(nèi)皮細(xì)胞和平滑肌細(xì)胞 / Pulmonary artery endothelial and smooth muscle cells	Collagenase: 0.04-0.05%； Soybean Trypsin Inhibitor: 0.04%	RPMI-1640	Yu, M., McAndrew, R., Al-Saghir, R., Maier, K., Medhora, M., Roman, R. and Jacobs, E.: Nitric Oxide Contributes to 20-HETE-Induced Relaxation of Pulmonary Arteries., J Appl Physiol 93, 1391, 2002
牛	Bovine	平滑肌 / Smooth muscle	Trypsin: 0.25%	DMEM	Absher, M., Woodcock-Mitchell, J., Mitchell, J., Baldor, L., Low, R., and Warshaw, D.: Characterization of Vascular Smooth Muscle Cell Phenotype in Long-Term Culture, In Vitro Cell Dev Biol 25 (2), 183, 1989
牛	Bovine	血管平滑肌 / Vascular smooth muscle	Elastase Type 3: 50 u/ml	PSS	Warshaw, D., Szarek, J., Hubbard, M., and Evans, J.: Pharmacology and Force Development of Single Freshly Isolated Bovine Cartoid Artery Smooth Muscle Cells, Circ Res 58, 399, 1986
牛	Bovine	平滑肌，成纖維細(xì)胞 / Smooth muscle, fibroblasts	Trypsin: 0.055%	DMEM	Davies, P. and Kerr, C.: Modification of LDL Metabolism by Growth Factors in Cultured Vascular Cells and Human Skin Fibroblasts, Biochim Biophys Acta 712, 26, 1982
犬/狗	Canine	骨骼肌 / Skeletal muscle	Collagenase Type 4: 200 u/ml； Neutral Protease: 1 u/ml	DMEM	Parker, M., Loretz, C., Tyler, A., Snider, L., Storb. R. and Tapscott, S.: Inhibition of CD26/DPP-IV Enhances Donor Muscle Cell Engraftment and Stimulates Sustained Donor Cell Proliferation., Skelet Muscle 2, 4, 2012
犬/狗	Dog	平滑肌 / Smooth muscle	Elastase: 50 u/ml	PSS	Subramanian, M., Madden, J., and Harder, D.: A Method for the Isolation of Cells from Arteries of Various Sizes, J Tiss Cul Meth 13, 13, 1991
犬/狗	Dog, beagle, adult	平滑肌，血管 / Smooth muscle，Vascular	Elastase: 34 u/ml	Tyrode's solution w/ calcium	Wilde, D., and Lee, K.: Outward Potassium Currents in Freshly Isolated Smooth Muscle Cell of Dog Coronary Arteries, Circ Res 65, 1718, 1989
犬/狗	Dog	動脈，頸動脈 / Artery，Carotid	Elastase: 80 u/ml	PSS	Dobrin, P., and Canfield, T.: Elastase, Collagenase, and the Biaxial Elastic Properties of Dog Carotid Artery, Am J Physiol Heart Circ Physiol 247 (16), H124, 1984
雞	Chicken, 1-2 day	胗和主動脈平滑肌 / Gizzard and aorta smooth muscle	Collagenase Type 1: 0.15%	HBSS	Dirksen W., Vladic F., and Fisher S.: A Myosin Phosphatase Targeting Subunit Isoform Transition Defines a Smooth Muscle Developmental Phenotypic Switch, Am J Physiol/Cell 278(3), C589, 2000
雞	Chick	平滑肌 / Smooth muscle	Trypsin: 0.05% - 0.1%	HBSS	Chamley-Campbell, J., Campbell, G., and Ross, R.: The Smooth Muscle in Cell Culture, Physiol Res 59, 1, 1979
雞	Chick, white leghorn, 12 day	肌肉 / Muscle	Trypsin: 0.25%	Puck's saline A	Bullaro, J., and Brookman, D.: Comparison of Skeletal Muscle Monolayer Cultures Initiated With Cells Dissociated by the Vortex and Trypsin Methods, In Vitro 12, 564, 1976
雞	Chick, white leghorn, embryos, 11 day	肌肉 / Muscle	Trypsin: 0.05%	Saline G	Tepperman, K., Morris, G., Essien, F., and Heywood, S.M.: A Mechanical Dissociation Method For Preparation of Muscle Cell Cultures, J Cell Physiol 86, 561, 1975
雞	Chick embryo	甲狀腺，肌肉，心臟 / Thyroid，Muscle，Heart	Collagenase: 0.25%	Tyrode's saline, potassium free	Hilfer, S., and Brown, J.: Collagenase. Its Effectiveness as a Dispersing Agent for Embryonic Chick Thyroid and Heart, Exp Cell Res 65, 246, 1971
雞	Chick, embryonic	肌肉 / Muscle	Trypsin: 0.1%	CMF HBSS	Hilfer, S.: Collagenase Treatment of Chick Heart and Thyroid,Tissue Cult Methods & Applications,Kruse, P., and Patterson, M., , 246, 1971
雞	Chick embryonic	各種組織（心臟，肝臟，骨骼，心臟） / Various tissues (heart, liver, skeletal, cardiac)	Trypsin: various grades	CMF Tyrode's solution	Rinaldini, L.: An Improved Method for the Isolation and Quantitative Cultivation of Embryonic Cells, Exp Cell Res 16, 477, 1959
貓	Cat, adult mongrel, either sex, 2.5-4.0 kg	腦動脈 / Cerebral arteries	Elastase: 50 u/ml	Puck's solution	Madden, J., Vadula, M., and Kurup, V.: Effects of Hypoxia and Other Vasoactive Agents on Pulmonary and Cerebral Artery Smooth Muscle Cells, Am J Physiol 263, L384, 1992
貓	Cat, mongrel, adult, 2-4 kg	肌細(xì)胞 / Myocytes	Collagenase: 0.12%	Kreb's Henseleit, CF	Follmer, C.H., Ten Eick, R.E., and Yeh, J.Z.: Sodium Current Kinetics in Cat Atrial Myocytes, J Physiol 384, 169, 1987
魚	Zebrafish	生肌祖細(xì)胞 / Myogenic progenitor	Collagenase Type 4: 0.5%； Neutral Protease: 1.2 u/ml	PBS	Alexander, M., Kawahara, G., Kho, A., Howell, M., Pusack, T., Myers, J., Montanaro, F., Zon, L., Guyon, J. and Kunkel, L.: Isolation and Transcriptome Analysis of Adult Zebrafish Cells Enriched for Skeletal Muscle Progenitors., Muscle Nerve 43, 741-50, 2011
魚	Dogfish, Squalus acanthias , 2-6 Kg	肌細(xì)胞，心臟和胃 / Myocytes, heart and stomach	Protease XIV: 0.028%	Solution C	Mitra, R. and Morad, M.: A Uniform Enzymatic Method for Dissociation of Myocytes from Hearts and Stomachs of Vertebrates, Am J Physiol 249, H1056, 1985
青蛙	Frog, Xenopus laevis, embryos stage 17 & 19	肌肉 / Muscle	Collagenase: 0.10%	Steinberg's solution	Stollberg, J. and Fraser, S.: Acetylcholine Receptors and Con A-Binding Sites on Xenopus Muscle Cells, J Cell Biol 107, 1397, 1988
青蛙	Frog	肌細(xì)胞 / Myocytes	Trypsin: 0.1%	CF Ringer	Shepherd, N. and Kavaler, F.: Direct Control of Contraction Force of Single Frog Atrial Cells by Extracellular Ions, Am J Physiol 251, C653, 1986
青蛙	Frog, Rana pipiens (50-100 g)	肌細(xì)胞，心臟和胃 / Myocytes, heart and stomach	Protease XIV: 0.028%	Solution C	Mitra, R. and Morad, M.: A Uniform Enzymatic Method for Dissociation of Myocytes from Hearts and Stomachs of Vertebrates, Am J Physiol 249, H1056, 1985
青蛙	Frog, Xenopus laevis	肌肉 / Muscle	Trypsin: 0.5%	L15 medium (see reference)	Anderson, M.J., Cohen, M.W., and Zorychta, E.: Effects of Innervation on the Distribution of Acetylcholine Receptors on Cultured Muscle Cells, J Physiol 268, 731, 1977
豚鼠	Guinea-pig	膀胱平滑肌 / Bladder smooth muscle	Collagenase Type 2: 0.1-0.2%	Krebs-Ringer bicarbonate	Shieh CC, Feng J, Buckner SA, Brioni JD, Coghlan MJ, Sullivan JP, Gopalakrishnan M: Functional implication of spare ATP-sensitive K(+) channels in bladder smooth muscle cells, J Pharmacol Exp Ther 296, 669-75, 2001
豚鼠	Guinea-pig, 200-380 g	毛細(xì)血管，肌細(xì)胞 / Capillaries，Myocytes	Collagenase Type 2: 0.15%	CF solution	Schnitzler, M., Derst, C., Daut, J., and Preisig-Muller, R.: ATP-Sensitive Potassium Channels in Capillaries Isolated From Guinea-Pig Heart, J Physiol 525 (2), 307, 2000
豚鼠	Guinea pig, 2-4 wk old, male, female	平滑肌，膽囊 / Smooth muscle，Gallbladder	Papain: 0.1%	Krebs solution	Firth, T., Mawe, G., and Nelson, M.: Pharmacology and Modulation of KATP Channels by Protein Kinase C and Phosphatates in Gallbladder Smooth Muscle, Am J Physiol Cell Physiol 278, C1031, 2000
豚鼠	Guinea-pig, adult, 250-350 g	平滑肌，膽囊 / Smooth muscle，Gallbladder	Papain: 0.1%	NaCl, sodium glutamate, MgCl, KCl, glucose, Kreb's, and HEPES	Jennings, L., Xu, Q., Firth, T., Nelson, M., and Mawe, G.: Cholesterol Inhibits Spontaneous Action Potentials and Calcium Currents in Guinea Pig Gallbladder Smooth Muscle, Am J Physiol 277, G1017, 1999
豚鼠	Guinea-pig, Dunkin-Hartley, female	肌細(xì)胞 / Myocytes	Protease:	DMEM	Ryder, K., Bryant, S., and Hart, G.: Membrane Current Changes in Left Ventricular Myocytes Isolated From Guinea-Pigs After Abdominal Aortic Coarctation, Cardiovasc Res 27, 1278, 1993
豚鼠	Guinea-pig (also rat, rabbit)	平滑肌 / Smooth muscle	Trypsin: 0.1%	Potassium buffer solution	Hu, S., and Kim, H.: Activation of K+ Channel in Vascular Smooth Muscle by Cytochrome P-450 Metabolites of Arachidonic Acid, FASEB J 6, A383, 1992
豚鼠	Guinea-pig, 200-300 g	平滑肌，腸系膜動脈 / Smooth muscle，Mesenteric artery	Collagenase: 0.3%	CF solution	Ohya, Y. and Sperelakis, N.: ATP Regulation of the Slow Calcium Channels in Vascular Smooth Muscle Cells, Circ Res 64, 145, 1989
豚鼠	Guinea-pig, 200-400 g	肌細(xì)胞，心臟和胃 / Myocytes, heart and stomach	Protease XIV: 0.028%	Solution C	Mitra, R. and Morad, M.: A Uniform Enzymatic Method for Dissociation of Myocytes from Hearts and Stomachs of Vertebrates, Am J Physiol 249, H1056, 1985
豚鼠	Guinea-pig, prepubertal	平滑肌 / Smooth muscle	Trypsin: 0.05%	Dulbecco-Vogt modification of Eagle's	Ross, R.: The Smooth Muscle Cell . II. Growth of Smooth Muscle in Culture and Formation of Elastic Fibers, J Cell Biol 50, 172, 1971
倉鼠	Hamster, male, 60-70 day	衛(wèi)星 / Satellite	Trypsin: 0.25%	DMEM	Nakamura, T., Iwata, Y., Sampaolesi, M., Hanada, H., Saito, N., Artman, M., Coetzee, W., and Shigekawa, M.: Stretch-Activated Cation Channels in Skeletal Muscle Myotubes From Sarcoglycan-Deficient Hamsters, Am J Physiol Cell Physiol 281, C690, 2001
人	Human	膀胱平滑肌 / Bladder smooth muscle	Collagenase Type 2: 0.1-0.2%； Papain: 0.1-0.2%	Digestion solution (see reference)	Malysz, J., Rovner, E., Wake, R. and Petkov, G.: Preparation and Utilization of Freshly Isolated Human Detrusor Smooth Muscle Cells for Characterization of 9-Phenanthrol-Sensitive Cation Currents., J Vis Exp , , 2020
人	Human	肌肉衛(wèi)星 / Muscle satellite	Collagenase: 0.1%； Trypsin: 0.25%	DMEM	Garcia, S., Tamaki, S., Lee, S., Wong, A., Jose, A., Dreux, J., Kouklis, G., Sbitany, H., Seth, R., Knott, P., Heaton, C., Ryan, W., Kim, E., Hansen, S., Hoffman, W. and Pomerantz, J.: High-Yield Purification, Preservation, and Serial Transplantation of Human Satellite Cells., Stem Cell Reports 10, 1160-1174, 2018