原代细胞分离中，针对肌肉组织涉及到肌肉卫星细胞，肺动脉内皮细胞和平滑肌细胞、平滑肌、血管平滑肌等等的解离通常会使用酶，如何选择合适的酶来完成对应肌肉组织的解离，以及原代细胞采集的常规操作， 艾美捷针对肌肉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：
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。
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| 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 | 肌肉卫星细胞 / 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 | 肺动脉内皮细胞和平滑肌细胞 / 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，成纤维细胞 / 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 | 动脉，颈动脉 / 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 |
| 鸡 | Chicken, 1-2 day | 胗和主动脉平滑肌 / 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 |
| 鸡 | 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 |
| 鸡 | 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 |
| 鸡 | 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 |
| 鸡 | Chick embryo | 甲状腺，肌肉，心脏 / 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 |
| 鸡 | 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 |
| 鸡 | Chick embryonic | 各种组织（心脏，肝脏，骨骼，心脏） / 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 |
| 猫 | Cat, adult mongrel, either sex, 2.5-4.0 kg | 脑动脉 / 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 |
| 猫 | Cat, mongrel, adult, 2-4 kg | 肌细胞 / 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 |
| 鱼 | Zebrafish | 生肌祖细胞 / 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 |
| 鱼 | Dogfish, Squalus acanthias , 2-6 Kg | 肌细胞，心脏和胃 / 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 | 肌细胞 / 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) | 肌细胞，心脏和胃 / 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 | 毛细血管，肌细胞 / 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，胆囊 / 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，胆囊 / 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 | 肌细胞 / 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，肠系膜动脉 / 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 | 肌细胞，心脏和胃 / 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 |
| 仓鼠 | Hamster, male, 60-70 day | 卫星 / 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 | 肌肉卫星 / 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 |
| 人 | Human | 骨骼肌干细胞 / Skeletal muscle stem | Collagenase Type 2: 750 u/ml； Neutral Protease: 2 u/ml |
Ham's F10 | Charville, G., Cheung, T., Yoo, B., Santos, P., Lee, G., Shrager, J. and Rando, T.: Ex Vivo Expansion and In Vivo Self-Renewal of Human Muscle Stem Cells., Stem Cell Reports 5, 621, 2015 |
| 人 | Human | 平滑肌 / Smooth muscle | Collagenase Type 1: 0.1% | DMEM | Lu, S., Sun, X., Hong, T., Song, K., Yang, S. and Wang, C.: Isolation and Culture of Smooth Muscle Cells from Human Acute Type A Aortic Dissection., J Cardiothorac Surg 8, 83, 2013 |
| 人 | Human, male | 肌源性 / Myogenic | Collagenase Type 4: 0.1%； Neutral Protease: 2.4 u/ml |
HBSS | Stadler, G., Chen, J., Wagner, K., Robin, J., Shay, J., Emerson, C. and Wright, W.: Establishment of Clonal Myogenic Cell Lines from Severely Affected Dystrophic Muscles - CDK4 Maintains the Myogenic Population., Skelet Muscle 1, 12, 2011 |
| 人 | Human | 肌肉来源的的多祖细胞 / Muscle derived multiprogenitor cells | Collagenase Type 2: 0.05% | DMEM | Nesti, L., Jackson, W., Shanti, R., Koehler, S., Aragon, A., Bailey, J., Sracic, M., Freedman, B., Giuliani, J. and Tuan, R.: Differentiation Potential of Multipotent Progenitor Cells Derived from War-Traumatized Muscle Tissue., J Bone Joint Surg Am 90, 2390, 2008 |
| 人 | Human | 内皮和血管平滑肌 / Endothelial and vascular smooth muscle | Collagenase Type 1: 0.2% | HBSS | Moss, S., Bates, M., Parrino, P. and Woods, TC.: Isolation of Endothelial Cells and Vascular Smooth Muscle Cells from Internal Mammary Artery Tissue., Ochsner J 7, 133, 2007 |
| 人 | Human | 泌尿道平滑肌 / Urinary tract smooth muscle | Collagenase Type 4: 100 u/ml | DMEM | Kimuli, M., Eardley, I. and Southgate, J.: In Vitro Assessment of Decellularized Porcine Dermis as a Matrix for Urinary Tract Reconstruction., BJU Int 94, 859, 2004 |
| 人 | Human, female | 平滑肌，肌层 / Smooth muscle，Myometrial | Deoxyribonuclease I: 0.015% and 0.007% | HBSS | Richardson, M., Taylor, D., Casey, M., MacDonald, P., and Stull, J.: Biochemical Markers of Contraction in Human Myometrial Smooth Muscle Cells in Culture, In Vitro Cell Dev Biol 23, 21, 1987 |
| 人 | Human, female | 平滑肌，肌层 / Smooth muscle，Myometrial | Deoxyribonuclease I: 0.12% | HBSS | Casey, M., MacDonald, P., Mitchell, M., and Snyder, J.: Maintenance and Characterization of Human Myometrial Smooth Muscle Cells in Monolayer Culture, In Vitro 20, 396, 1984 |
| 人 | Human, fetal | 平滑肌，成纤维细胞 / 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 |
| 人 | Human | 平滑肌 / Smooth muscle | Trypsin: 0.25% | DMEM | Eskin, S., Sybers, H., Lester, J., Navarro, L., Gotto, A., and DeBakey, M.: Human Smooth Muscle Cells Cultured From Atherosclerotic Plaques and Uninvolved Vessel Wall, In Vitro 17 (8), 713, 1981 |
| 人 | Human | 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 |
| 蜥蜴 | Lizard (Anolis carolinensis) | 成肌细胞，尾巴 / Myoblasts, tail | Collagenase: 0.2% | GM III | Cox, P., and Simpson, Jr., S.: A Microphotometric STudy of Myogenic Lizard Cells Grown in Vitro, Dev Biol 23, 433, 1970 |
| 猴 | Monkey (Macaca nemestrina) | 平滑肌 / Smooth muscle | Trypsin: 0.05% | Dulbecco-Vogt | Chait, A., Ross, R., Albers, J., and Bierman, E.: Platelet-Derived Growth Factor Stimulates Activity of LDL Receptors, Proc Natl Acad Sci U S A 77, 4084, 1980 |
| 猴 | Rhesus monkey, 1 year | 平滑肌，大隐静脉 / Smooth muscle, saphenous vein | Elastase: 0.05% | BSS | Chamley, J., Campbell, G., McConnell, J., and Groschel-Stewart, U.: Comparison of Vascular Smooth Muscle Cells from Adult Human, Monkey and Rabbit in Primary Culture and in Subculture, Cell Tissue Res 177, 503, 1977 |
| 小鼠 | 肌纤维 / Myofiber | Collagenase Type 1: 0.2% | DMEM | Huttner,S., Ahrens, H., Schmidt, M., Henze, H., Jung, M., Schuler. S. and von Maltzahn, J.: Isolation and Culture of Individual Myofibers and Their Adjacent Muscle Stem Cells from Aged and Adult Skeletal Muscle., Methods Mol Biol , , 2019 | |
| 小鼠 | Mouse | 眼外肌 / Extraocular muscles | Collagenase Type 1: 0.1%； Collagenase Type 2: 0.03-0.2%； Elastase: 0.0125-0.1% |
Ringer | Eckhardt, J., Bachmann, C., Sekulic-Jablanovic, M., Enzmann, V., Park, K., Ma, J., Takeshima, H., Zorzato, F. and Treves, S.: Extraocular Muscle Function is Impaired in ryr3−/− Mice, J Gen Physiol 151, 929-943, 2019 |
| 小鼠 | Mouse, 6-8 week | 血管平滑肌 / vascular smooth muscle | Collagenase Type 1: 0.1%； Elastase: 0.75 u/ml |
DMEM/F12 | Sachdeva, J., Mahajan, A., Cheng, J., Baeten, J., Lilly, B., Kuivaniemi, H. and Hans, C.: Smooth Muscle Cell-Specific Notch1 Haploinsufficiency Restricts the Progression of Abdominal Aortic Aneurysm by Modulating CTGF Expression., PLoS ONE 12, e0178538, 2017 |
| 小鼠 | Mouse, GFP transgenic | 肌肉来源的的 NLSC / Muscle derived NLSC | Collagenase Type 2: 0.2% | DMEM | Birbrair, A, Sattiraju, A, Zhu, D, Zulato, G, Batista, I, Nguyen, V, Messi M, Solingapuram Sai, K, Marini, F, Delbono, O and Mintz, A.: Novel Peripherally Derived Neural-Like Stem Cells as Therapeutic Carriers for Treating Glioblastomas., Stem Cells Transl Med , , 2016 |
| 小鼠 | Mouse, 2-4 month | 咽肌纤维 / Pharyngeal myofiber | Collagenase Type 1: 400-800 u/ml | DMEM | Randolph, M., Phillips, B., Choo, H., Vest, K., Vera, Y. and Pavlath, G.: Pharyngeal Satellite Cells Undergo Myogenesis Under Basal Conditions and Are Required for Pharyngeal Muscle Maintenance., Stem Cells 33, 3581-95, 2015 |
| 小鼠 | Mouse, 2 month | 成肌细胞 / Myoblasts | Collagenase Type 2: 400 u/ml； Neutral Protease: 0.5 u/ml |
Hams F-10 | Abou-Khalil, R., Yang, F., Lieu, S., Julien, A., Perry, J., Pereira, C., Relaix, F., Miclau, T., Marcucio, R. and Colnot, C.: Role of Muscle Stem Cells During Skeletal Regeneration., Stem Cells 33, 1501-11, 2015 |
| 小鼠 | Mouse, 6-8 week | 肌纤维 / Myofibers | Collagenase Type 1: 0.2% | DMEM | Pasut, A., Jones, A. and Rudnicki, M.: Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle., J Vis Exp 73, e50074, 2013 |
| 小鼠 | Mouse | 血管平滑肌 / Vascular smooth muscle | Collagenase Type 2: 0.14% | Basal Culture | Metz, R., Patterson, J. and Wilson, E.: Vascular Smooth Muscle Cells: Isolation, Culture, and Characterization., Methods Mol Biol 843, 169, 2012 |
| 小鼠 | Mouse, 10 week | 隔膜细胞 / Diaphragm cells | Collagenase Type 2: 100 u/ml； Pronase: 0.125% |
PBS | Rezk, B., Yoshida, T., Semprun-Prieto, L., Higashi, Y., Sukhanov, S. and Delafontaine, P.: Angiotensin II Infusion Induces Marked Diaphragmatic Skeletal Muscle Atrophy., PLoS ONE 7, e30276, 2012 |
| 小鼠 | Mouse | 骨骼肌纤维 / Skeletal muscle fiber | Collagenase Type 1: 0.2% | Tyrode | Weisleder, N., Zhou, J. and Ma, J.: Detection of Calcium Sparks in Intact and Permeabilized Skeletal Muscle Fibers., Methods Mol Biol 798, 395, 2012 |
| 小鼠 | Mouse | 成肌细胞 / Myoblast | Collagenase Type 2: 0.2% | DMEM | Shi, H., Boadu, E., Mercan, F., Le, A., Roth Flach, R., Zhang, L., Tyner, K., Olwin, B. and Bennett, A.: MAP Kinase Phosphatase-1 Deficiency Impairs Skeletal Muscle Regeneration and Exacerbates Muscular Dystrophy., FASEB J 24, 2985, 2010 |
| 小鼠 | Mouse, 1-25 month | 肌细胞，内皮细胞 / Myocytes, endothelial | Neutral Protease: 1.2 u/ml； Collagenase Type 4: 0.2% |
PBS | Ieronimakis Nicholas, Balasundaram Gayathri, Reyes Morayma: Direct isolation, culture and transplant of mouse skeletal muscle derived endothelial cells with angiogenic potential, PLoS ONE 3, e0001753, 2008 |
| 小鼠 | Mouse, 15-20 day | 卡哈尔间质细胞 / Intersitial cells of Cajal | Collagenase Type 2: 0.13% | M199 | Li CX, Liu BH, Tong WD, Zhang LY, and Jiang YP: Dissociation, culture and morphologic changes of interstitial cells of Cajal in vitro, World J Gastroenterol 11, 2838, 2005 |
| 小鼠 | Mouse, neonatal | 骨骼肌肌管 / Skeletal muscle myotubes | NCIS kit: per instructions | L-15 | Johnson, B., Scheuer, T. and Catterall, W.: Convergent Regulation of Skeletal Muscle Ca2+ Channels by Dystrophin, the Actin Cytoskeleton, and cAMP-Dependent Protein Kinase., Proc Natl Acad Sci U S A 102, 4191, 2005 |
| 小鼠 | Mouse, male, 6-14 week | 前体细胞 / Precursor cells | Collagenase Type 2: 0.5% | DMEM/F12 | Winitsky, S., Gopal, T., Hassanzadeh, S., Takahashi, H., Gryder, D., Rogawski, M., Takeda, K., Yu, Z., Xu, Y. and Epstein, N.: Adult Murine Skeletal Muscle Contains Cells That Can Differentiate into Beating Cardiomyocytes In Vitro., PLoS Biol 3, e87, 2005 |
| 小鼠 | Mouse, 6-8 week | 骨骼肌祖细胞 / Skeletal muscle progenitor | Collagenase Type 2: 0.2% | DMEM | Majka S., Jackson K., Kienstra K., Majesky M., Goodell M., and Hirschi K.: Distinct Progenitor Populations in Skeletal Muscle Are Bone Marrow Derived and Exhibit Different Cell Fates During Vascular Regeneration, J Clin Invest 111, 71, 2003 |
| 小鼠 | Mouse, neonatal | 肌细胞 / Myocytes | Collagenase Type 1: 0.5% | DMEM | Fukada S., Miyagoe-Suzuki Y., Tsukihara H., Yuasa K., Higuchi S., Ono S., Tsujikawa K., Takeda S., and Yamamoto H.: Muscle Regeneration by Reconstitution with Bone Marrow or Fetal Liver Cells from Green Fluorescent Protein-gene Transgenic Mice, J Cell Sci 115, 1285, 2002 |
| 小鼠 | Mouse, 6-8 week | 肌细胞 / Myocytes | Collagenase Type 2: 0.2%； Trypsin: 0.25% |
HBSS | McKinney-Freeman SL, Jackson KA, Camargo FD, Ferrari G, Mavilio F, Goodell MA.: Muscle-derived hematopoietic stem cells are hematopoietic in origin, Proc Natl Acad Sci U S A 99, 1341, 2002 |
| 绵羊 | Sheep, adult and neonatal | 气管平滑肌细胞 / Tracheal smooth muscle cells | Papain: 0.2%； Deoxyribonuclease I: 0.1% |
MOPS-PSS | Driska S., Laudadio R., Wolfson M., and Shaffer T.: A Method for Isolating Adult and Neonatal Airway Smooth Muscle Cells and Measuring Shortening Velocity, J Appl Physiol 86(1), 427, 1999 |
| 猪 | Porcine, postnatal | 肌肉卫星 / Muscle satellite | Trypsin: 0.25%； Collagenase Type 1: 0.2%； Deoxyribonuclease I: 0.01% |
Eagle MEM | Miersch, C., Stange, K. and Rontgen, M.: Effects of Trypsinization and of a Combined Trypsin, Collagenase, and DNase Digestion on Liberation and in Vitro Function of Satellite Cells Isolated from Juvenile Porcine Muscles., In Vitro Cell Dev Biol Anim 54, 406-412, 2018 |
| 猪 | Porcine, 2 month | 骨骼肌 / Skeletal muscle | Collagenase: 10%； Neutral Protease: 0.3% |
HBSS | Lewis, F., Henning, B., Marazzi, G., Sassoon, D., Ellison, G. and Nadal-Ginard, B.: Porcine Skeletal Muscle-Derived Multipotent PW1pos/Pax7neg Interstitial Cells: Isolation, Characterization, and Long-Term Culture., Stem Cells Transl Med 3, 702-12, 2014 |
| 猪 | Porcine, male, 25-40 kg | 冠状动脉肌细胞 / Coronary myocytes | Collagenase Type 2: 294 u/ml； Elastase: 6.5 u/ml； Deoxyribonuclease I: 0.04%； Soybean Trypsin Inhibitor: 0.1% |
low calcium physiological saline | Korzick, D., Laughlin, M., and Bowles, D.: Alterations in PKC signaling underlie enhanced myogenic tone in exercise-trained porcine coronary resistance arteries, J Appl Physiol 96, 1425-32, 2004 |
| 猪 | Porcine, adult, 35-45 kg | 动脉平滑肌 / Arterial smooth muscle | Collagenase Type 2: 294 u/ml； Elastase: 6.5 u/ml； Deoxyribonuclease I: 0.4 mg/ml； Soybean Trypsin Inhibitor: 1 mg/ml |
MEM | Wamhoff BR, Dixon JL, Sturek M: Atorvastatin treatment prevents alterations in coronary smooth muscle nuclear Ca2+ signaling in diabetic dyslipidemia, J Vasc Res 39, 208, 2002 |
| 猪 | Porcine, Yorkshire, 30kg | 冠状平滑肌 / Coronary smooth muscle | Collagenase Type 2: 150 u/ml； Elastase: 0.05% |
HBSS | Sirous ZN, Fleming JB, Khalil RA: Endothelin-1 enhances eicosanoids-induced coronary smooth muscle contraction by activating specific protein kinase C isoforms, Hypertension 37, 497-504, 2001 |
| 猪 | Porcine | 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 |
| 猪 | Porcine, juvenile, 30kg | 冠状动脉平滑肌细胞 / Coronary smooth muscle cells | Collagenase: 0.3%； Elastase: 0.05% |
HBSS | Huckle WR, Drag MD, Acker WR, Powers M, McFall RC, Holder DJ, Fujita T, Stabilito II, Kim D, Ondeyka DL, Mantlo NB, Chang RS, Reilly CF, Schwartz RS, Greenlee WJ, Johnson RG: Effects of subtype-selective and balanced angiotensin II receptor antagonists in a porcine coronary artery model of vascular restenosis, Circulation 93, 1009-19, 1996 |
| 猪 | Porcine | 平滑肌，主动脉 / Smooth muscle, aorta | Collagenase: 0.3% | DMEM | Xiong, Yimin, Xu, Shangzhe, and Slakey, Linda L: Modulation of Response to Adenosine in Vascular Smooth Muscle Cells Cultured in Defined Medium, In Vitro Cell Dev Biol 27, 355, 1991 |
| 猪 | Porcine | 平滑肌 / Smooth muscle | Trypsin: 0.05% | EDTA 0.02% | Breton, M., Berrou, E., Deudon, E. and Picard, J.: Changes in Proteoglycans of Cultured Pig Aortic Smooth Muscle Cells During Subculture, In Vitro Cell Dev Biol 26, 157, 1990 |
| 猪 | Porcine | 平滑肌 / Smooth muscle | Collagenase: 0.30% | DMEM | Fehr, T., Dickinson, E., Goldman, S. and Slakey, L.: Cyclic AMP Efflux is Regulated by Occupancy of The Adenosine Receptor in Pig Aortic Smooth Muscle Cells, J Biol Chem 265, 10974, 1990 |
| 鹌鹑 | Quail, embryo, 10 day | 成肌细胞 / Myoblasts | Collagenase Type 2: 0.1% | Puck's solution | Konigsberg, I.: Skeletal Myoblasts in Culture, Methods Enzymol LVIII, 511, 1979 |
| 兔 | Rabbit, New Zealand | 胃平滑肌 / Gastric smooth muscle | Collagenase Type 2: 0.1%； Soybean Trypsin Inhibitor: 0.01% |
DMEM | Al-Shboul, O., Mahavadi, S., Sriwai, W., Grider, J. and Murthy, K.: Differential Expression of Multidrug Resistance Protein 5 and Phosphodiesterase 5 and Regulation of cGMP Levels in Phasic and Tonic Smooth Muscle., Am J Physiol Gastrointest Liver Physiol 305, G314-24, 2013 |
| 兔 | Rabbit, New Zealand White | 主动脉平滑肌 / Aortic smooth muscle | Collagenase Type 2: 300 u/ml； Elastase: 5 u/ml |
F10 Ham's | Croons Valerie, Martinet Wim, Herman Arnold G, Timmermans Jean-Pierre, De Meyer Guido R Y: Selective clearance of macrophages in atherosclerotic plaques by the protein synthesis inhibitor cycloheximide, J Pharmacol Exp Ther 320, 986-93, 2007 |
| 兔 | Rabbit, New Zealand, male, 2.5-3.5 Kg | 肠细胞 / Enterocytes | Trypsin: 0.1% | RPMI 1640 w/ 1% fetal bovine serum；PBS | Santos, M., Nguyen, B.., Thompson, J.: Factors Affecting in Vitro Growth of Harvested Enterocytes, Cell Transplant 1, 299, 1992 |
| 兔 | 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 |
| 兔 | Rabbit, adult, 1-2 kg | 平滑肌 / Smooth muscle | Elastase: 0.17 - 0.25% | Saline | Benham, C., Bolton, T., Byrne, N., and Large, W.: Action of Extremely Applied Adenosine Triphosphate O Single Smooth Muscle Cells Dispersed From Rabbit Ear Artery, J Physiol 387, 473, 1987 |
| 兔 | Rabbit, New Zealand white, 1500 g | 平滑肌，主动脉 / Smooth muscle, aortic | Trypsin: 0.038% | MEM | Knodle, S., Anderson, S., and Papaioannou, S.: Large Scale Preparation of Rabbit Aortic Smooth Muscle Cells For Use in Calcium Uptake Studies, In Vitro Cell Dev Biol 22, 23, 1986 |
| 兔 | Rabbit, adult, 1-2 Kg | 平滑肌，耳动脉 / Smooth muscle, ear artery | Trypsin: 0.1% | CF solution | Benham, C.D., Bolton, T.B.: Spontaneous Transient Outward Currents in Single Visceral and Vascular Smooth Muscle Cells of the Rabbit, J Physiol 381, 385, 1986 |
| 兔 | Rabbit, 0.5-1 Kg | 肌细胞，心脏和胃 / 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 |
| 兔 | Rabbit, white New Zealand, adult, male, 2 Kg | 平滑肌，主动脉 / Smooth muscle, aorta | Trypsin: 0.1% | Krebs Ringer HEPES solution | Ives, H., Schultz, G., Galardy, R., and Jamieson, J.: Preparation of Functional Smooth Muscle Cells from the Rabbit Aorta, J Exp Med 148, 1400, 1978 |
| 兔 | Rabbits, New Zealand white albino, 5-6 months | 平滑肌，大隐静脉 / Smooth muscle, saphenous vein | Elastase: 0.05% | BSS | Chamley, J., Campbell, G., McConnell, J., and Groschel-Stewart, U.: Comparison of Vascular Smooth Muscle Cells from Adult Human, Monkey and Rabbit in Primary Culture and in Subculture, Cell Tissue Res 177, 503, 1977 |
| 兔 | Rabbit, chinchilla, 5-8 month, virgin, female, 2-3 kg | 平滑肌，主动脉 / Smooth muscle, aorta | Hyaluronidase: 800 u/ml | HBSS | Peters, T., Muller, M., and deDuve, C.: Lysosomes of the Arterial Wall. I. Isolation and Subcellular Fractionation of Cells from Normal Rabbit Aorta, J Exp Med 136, 1117, 1972 |
| 兔 | Rabbit, New England, albino | 胸主动脉 / Thoracic aorta | Elastase: 0.008% | Kreb's Ringer | Day, A., Phil, D., and Newman, H.: Synthesis of Phospholipid by Foam Cells Isolated from Rabbit Atherosclerotic Lesions, Circ Res XIX, 122, 1966 |
| 大鼠 | Rat, day 7 | 骨骼肌 / Skeletal muscle | Collagenase: 0.05%； Neutral Protease: 0.35% |
DMEM | Vilmont, V., Cadot, B., Ouanoumou, G. and Gomes, E.: A System for Studying Mechanisms of Neuromuscular Junction Development and Maintence, Development 143, 2464, 2016 |
| 大鼠 | Rat, SD | 肺动脉平滑肌 / Pulmonary arterial smooth muscle | Collagenase Type 2: 0.1%； Elastase: 0.05% |
Buffer | Zeng, Y., Liu, H., Kang, K., Wang, Z., Hui, G., Zhang, X., Zhong, J., Peng, W., Ramchandran, R., Raj, J. and Gou, D.: Hypoxia Inducible Factor-1 Mediates Expression of miR-322: Potential Role in Proliferation and Migration of Pulmonary Arterial Smooth Muscle Cells., Sci Rep 5, 12098, 2015 |
| 大鼠 | Rat | 骨骼肌纤维 / Skeletal muscle fiber | Collagenase Type 1: 0.2% | Tyrode | Weisleder, N., Zhou, J. and Ma, J.: Detection of Calcium Sparks in Intact and Permeabilized Skeletal Muscle Fibers., Methods Mol Biol 798, 395, 2012 |
| 大鼠 | Rat | 血管平滑肌 / Vascular smooth muscle | Collagenase Type 2: 0.1%； Elastase: 0.02%； Soybean Trypsin Inhibitor: 0.05%； Deoxyribonuclease I: 0.01% |
DMEM | Weber, S., Gratopp, A., Akanbi, S., Rheinlaender, C., Sallmon, H., Barikbin, P. and Koehne, P.: Isolation and Culture of Fibroblasts, Vascular Smooth Muscle, and Endothelial Cells From the Fetal Rat Ductus Arteriosus, Pediatr Res 70, 236, 2011 |
| 大鼠 | Rat, fetal 17-18 day | 肌管 / Myotubes | Trypsin: 0.05% | see reference | Das, M., Rumsey, J., Bhargava, N., Stancescu, M. and Hickman, J.: Skeletal Muscle Tissue Engineering: A Maturation Model Promoting Long-Term Survival of Myotubes, Structural Development of the Excitation-Contraction Coupling Apparatus and Neonatal Myosin Heavy Chain Expression., Biomaterials 30, 5392, 2009 |
| 大鼠 | Rat | 肌源性 / Myogenic | Collagenase Type 2: 1.0%； Neutral Protease: 2.4 u/ml |
Ham's F-10 | Pacak, C. and Cowan, D.: Fabrication of Myogenic Engineered Tissue Constructs., J Vis Exp 27, e1137, 2009 |
| 大鼠 | Rat | 肌源性 / myogenic | Neutral Protease: 8 u/ml； Collagenase Type 4: 200 u/ml |
DMEM | Larkin, L., Calve, S., Kostrominova, T. and Arruda, E.: Structure and Functional Evaluation of Tendon-Skeletal Muscle Constructs Engineered In Vitro., Tiss Eng 12, 3149-58, 2006 |
| 大鼠 | Rat, Lewis, neonatal | 成肌细胞 / Myoblasts | Collagenase Type 2: 1.0%； Neutral Protease: 2.4 u/ml |
Ham's F-10 | Kim, J., Hadlock, T., Cheney, M., Varvares, M. and Marler, J.: Muscle Tissue Engineering for Partial Glossectomy Defects., Arch Facial Plast Surg 5, 403, 2003 |
| 大鼠 | Rat, SD, adult and neonatal w/in 1 day of birth | 肌样 / Myooids | Neutral Protease: 4 u/ml | Ham's F-12 | Dennis, R., Kosnik II, P., Gilbert, M., and Faulkner, J.: Excitability and Contractility of Skeletal Muscle Engineered from Primary Cultures and Cell Lines, Am J Physiol Cell Physiol 280, C288, 2001 |
| 大鼠 | Rat, SD, 3 month | 平滑肌细胞 / Smooth muscle cells | Collagenase Type 2: 0.2%； Elastase: 0.04%； Soybean Trypsin Inhibitor: 0.1% |
M-199 | Su E., Stevenson S., Rollence M., Marshall-Neff J., and Liau G.: A Genetically Modified Adenoviral Vector Exhibits Enhanced Gene Transfer of Human Smooth Muscle Cells, J Vasc Res 38(5), 471, 2001 |
| 大鼠 | Rat, SD, 250 g | 动脉平滑肌 / Arterial smooth muscle | Papain: 0.03%； Collagenase: 0.1% |
see reference | Jaggar JH: Intravascular pressure regulates local and global Ca(2+) signaling in cerebral artery smooth muscle cells, Am J Physiol Cell Physiol 281, C439-48, 2001 |
| 大鼠 | Rat, Wistar, adult, male | 平滑肌和骨骼肌 / Smooth & skeletal muscle | Protease: 0.01% | PSS | Wellman, G., Barrett-Jolley, R., Koppel, H. Everitt, D., and Quayle, J.: Inhibition of Vascular KATP Channels by U-37883A: A Comparison with Cardiac and Skeletal Muscle, Br J Pharmacol 128, 909, 1999 |
| 大鼠 | Rat, SD, male, 250-350 g | 血管平滑肌 / Vascular smooth muscle | Collagenase Type 2: 0.1%； Elastase: 0.0125% |
DMEM | Hrometz, S., Edelmann, S., McCune, D., Olges, J., Hadley, R., Perez ,D., and Piascik, M.: Expression of Multiple Alpha1-Adrenoceptors on Vascular Smooth Muscle: Correlation with the Regulation of Contraction, J Pharmacol Exp Ther 290(1), 452, 1999 |
| 大鼠 | Rat, SD, male, 150-175 g | 平滑肌，内皮 / Smooth muscle, endothelial | Trypsin: 0.04% | MEM | Redmond, E., Cahill, P., and Sitzmann, J.: Perfused Transcapillary Smooth Muscle and Endothelial Cell Co-Culture-A Novel In Vitro Model, In Vitro Cell Dev Biol Anim 31, 601, 1995 |
| 大鼠 | Rat, Wistar Kyoto, 10-15 weeks | 平滑肌，肠系膜动脉 / Smooth muscle, mesenteric artery | Trypsin: 0.05% | MEM | McGuire, P., Walker-Caprioglio, H., Little, S., and McGuffee, L.: Isolation and Culture of Rat Superior Mesenteric Artery Smooth Muscle Cells, In Vitro Cell Dev Biol 29, 135, 1993 |
| 大鼠 | Rat | 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 |
| 大鼠 | Rat, SD, 19 days | 平滑肌，肌层 / Smooth muscle, myometrial | Trypsin: 150 µg/ml | HBSS or PSS, CMF | Loch-Caruso, R., Pahl, M., and Juberg, D.: Rat Myometrial Smooth Muscle Cells Show High Levels of Gap Junctional Communication Under a Variety of Culture Conditions, In Vitro Cell Dev Biol 28, 97, 1992 |
| 大鼠 | Rat, SHRs and WKY, male, 10 - 14 weeks | 平滑肌，尾动脉 / Smooth muscle, tail arteries | Papain: 0.1% | HEPES buffer | Bolzon, B. and Cheung, D.: Isolation and Characterization of Single Vascular Smooth Muscle Cells From Spontaneously Hypertensive Rats, Hypertension 14, 137, 1989 |
| 大鼠 | Rat, 1-3 day | 平滑肌，主动脉 / Smooth muscle, aortic | Elastase: 0.0125% | DMEM | Barone, L., Wolfe, L., Faris, B., and Franzblau, C.: Elastin mRNA Levels and Insoluble Elastin Accumulation in Neonatal Rat, Biochemistry 27, 3175, 1988 |
| 大鼠 | Rat | 肌肉，肠系膜动脉 / Muscle, mesenteric arteries | Trypsin: 0.05% | HEPES KG solution | Bean, B., Sturek, M., Puga, A., and Hermsmeyer, K.: Calcium Channels in Muscle Cells Isolated From Rat Mesenteric Arteries: Modulation by Dihydropyridine Drugs, Circ Res 59, 229, 1986 |
| 大鼠 | Rat, SHR, WKY. either sex, 12-19 day, 3 month, and retired breeders | 内皮，主动脉 / Endothelial, aortic | Elastase: 0.05% | Waymouth's culture medium | Gordon, D., Mohai, L., and Schwartz, S.: Induction of Polyploidy in Cultures of Neonatal Rat Aortic Smooth Muscle Cells, Circ Res 59, 633, 1986 |
| 大鼠 | Rat, male, 150-250 g | 内皮，主动脉 / Endothelial, aortic | Trypsin: | RPMI 1640 | Cole, O., Fan, T., and Lewis, G.: Isolation, Characterization, Growth and Culture of Endothelial Cells From the Rat Aorta, Cell Biol Int Rep 10 (6), 399, 1986 |
| 大鼠 | Rat, Wistar, female, 10 weeks | 肌细胞 / Myocytes | Collagenase: 0.1% | HBSS | Boulanger-Saunier, C., Kattenburg, D., and Stoclet, J.: Cyclic AMP-dependent Phosphorylation of a 16kDa Protein in a Plasma Membrane-enriched Fraction of Rat Aortic Myocytes, F.E.B.S. Lett. 193, 283, 1985 |
| 大鼠 | Rat, SD, male, 200 - 250 g | 平滑肌，胸主动脉 / Smooth muscle, thoracic aorta | Trypsin: 0.0375% | Eagle's MEM with calcium | Brock, T., Alexander, R., Ekstein, L., Atkinson, W., and Gimbrone, M.: Angiotensin Increases Cytosolic Free Calcium in Cultured Vascular Smooth Muscle Cells, Hypertension 7, 105, 1985 |
| 大鼠 | Rat, SD, 225-250g | 肠系膜动脉平滑肌细胞 / Mesenteric artery smooth muscle cells | Elastase: .0125%； Soybean Trypsin Inhibitor: 0.025%； Collagenase Type 1: 0.1% |
HBSS | Gunther S, Alexander RW, Atkinson WJ, and Gimbrone MA Jr.: Functional angiotensin II receptors in cultured vascular smooth muscle cells, J Cell Biol 92, 289, 1982 |
| 大鼠 | Rat, 3-4 day | 心肌 / Myocardial | Trypsin NF 1:250: 0.125% | HBSS CMF | Kasten, F.: Rat Myocardial Cells In Vitro: Mitosis and Differentiated Properties, In Vitro 8, 128, 1972 |
| 大鼠 | Rat, 200 g | 肌肉 / Muscle | Trypsin: 0.05% | Kreb's Henseleit bicarbonate buffer | Kono, T.: Roles of Collagenases and Other Proteolytic Enzymes in the Dispersal of Animal Tissues, Biochim Biophys Acta 178, 397, 1969 |
| 大鼠 | Rat, Wistar, 3-10 day | 心脏 / Heart | Trypsin NF 1:250: 250: 0.1% | Saline A | Harary, I., and Farley, B.: In Vitro Studies on Single Beating Rat Heart Cells, Exp Cell Res 29, 451, 1963 |
