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worthington 肌肉组织分离指南

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:worthington肌肉组织分离指南

 

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

肌肉
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肌肉卫星细胞 / Muscle satellite cellsCollagenase Type 2: 300 u/mlKrebs-Ringer BicarbonateLee, 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
Bovine肺动脉内皮细胞和平滑肌细胞 / Pulmonary artery endothelial and smooth muscle cellsCollagenase: 0.04-0.05%;
Soybean Trypsin Inhibitor: 0.04%
RPMI-1640Yu, 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
Bovine平滑肌 / Smooth muscleTrypsin: 0.25%DMEMAbsher, 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
Bovine血管平滑肌 / Vascular smooth muscleElastase Type 3: 50 u/mlPSSWarshaw, 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
Bovine平滑肌,成纤维细胞 / Smooth muscle, fibroblastsTrypsin: 0.055%DMEMDavies, 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 muscleCollagenase Type 4: 200 u/ml;
Neutral Protease: 1 u/ml
DMEMParker, 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 muscleElastase: 50 u/mlPSSSubramanian, 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,VascularElastase: 34 u/mlTyrode's solution w/ calciumWilde, 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,CarotidElastase: 80 u/mlPSSDobrin, 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
Chicken, 1-2 day胗和主动脉平滑肌 / Gizzard and aorta smooth muscleCollagenase Type 1: 0.15%HBSSDirksen 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
Chick平滑肌 / Smooth muscleTrypsin: 0.05% - 0.1%HBSSChamley-Campbell, J., Campbell, G., and Ross, R.: The Smooth Muscle in Cell Culture, Physiol Res 59, 1, 1979
Chick, white leghorn, 12 day肌肉 / MuscleTrypsin: 0.25%Puck's saline ABullaro, 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
Chick, white leghorn, embryos, 11 day肌肉 / MuscleTrypsin: 0.05%Saline GTepperman, 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
Chick embryo甲状腺,肌肉,心脏 / Thyroid,Muscle,HeartCollagenase: 0.25%Tyrode's saline, potassium freeHilfer, S., and Brown, J.: Collagenase. Its Effectiveness as a Dispersing Agent for Embryonic Chick Thyroid and Heart, Exp Cell Res 65, 246, 1971
Chick, embryonic肌肉 / MuscleTrypsin: 0.1%CMF HBSSHilfer, S.: Collagenase Treatment of Chick Heart and Thyroid,Tissue Cult Methods & Applications,Kruse, P., and Patterson, M., , 246, 1971
Chick embryonic各种组织(心脏,肝脏,骨骼,心脏) / Various tissues (heart, liver, skeletal, cardiac)Trypsin: various gradesCMF Tyrode's solutionRinaldini, L.: An Improved Method for the Isolation and Quantitative Cultivation of Embryonic Cells, Exp Cell Res 16, 477, 1959
Cat, adult mongrel, either sex, 2.5-4.0 kg脑动脉 / Cerebral arteriesElastase: 50 u/mlPuck's solutionMadden, 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
Cat, mongrel, adult, 2-4 kg肌细胞 / MyocytesCollagenase: 0.12%Kreb's Henseleit, CFFollmer, C.H., Ten Eick, R.E., and Yeh, J.Z.: Sodium Current Kinetics in Cat Atrial Myocytes, J Physiol 384, 169, 1987
Zebrafish生肌祖细胞 / Myogenic progenitorCollagenase Type 4: 0.5%;
Neutral Protease: 1.2 u/ml
PBSAlexander, 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
Dogfish, Squalus acanthias , 2-6 Kg肌细胞,心脏和胃 / Myocytes, heart and stomachProtease XIV: 0.028%Solution CMitra, 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 / MuscleCollagenase: 0.10%Steinberg's solutionStollberg, 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 / MyocytesTrypsin: 0.1%CF RingerShepherd, 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 stomachProtease XIV: 0.028%Solution CMitra, 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 / MuscleTrypsin: 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 muscleCollagenase Type 2: 0.1-0.2%Krebs-Ringer bicarbonateShieh 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,MyocytesCollagenase Type 2: 0.15%CF solutionSchnitzler, 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,GallbladderPapain: 0.1%Krebs solutionFirth, 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,GallbladderPapain: 0.1%NaCl, sodium glutamate, MgCl, KCl, glucose, Kreb's, and HEPESJennings, 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 / MyocytesProtease:DMEMRyder, 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 muscleTrypsin: 0.1%Potassium buffer solutionHu, 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 arteryCollagenase: 0.3%CF solutionOhya, 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 stomachProtease XIV: 0.028%Solution CMitra, 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 muscleTrypsin: 0.05%Dulbecco-Vogt modification of Eagle'sRoss, 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 / SatelliteTrypsin: 0.25%DMEMNakamura, 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
Human膀胱平滑肌 / Bladder smooth muscleCollagenase 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
Human肌肉卫星 / Muscle satelliteCollagenase: 0.1%;
Trypsin: 0.25%
DMEMGarcia, 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


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