worthington 内皮细胞组织分离指南

发布者：艾美捷科技发布时间：2022-06-01

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原代细胞分离中，针对脑组织涉及到微血管内皮、脑内皮细胞、神经元和小胶质细胞、微血管等等的解离通常会使用酶，如何选择合适的酶来完成对应脑组织的解离，以及原代细胞采集的常规操作， 艾美捷针对脑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：

内皮组织分离指南

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。

内皮细胞
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	牛脐带 (BUVEC) / Bovine umbilical cord (BUVEC)	Collagenase: 0.1%	Dulbecco's/Ham F-12	Ricken Albert M, Traenkner Anja, Merkwitz Claudia, Hummitzsch Katja, Grosche Jens, Spanel-Borowski Katharina: The short prolactin receptor predominates in endothelial cells of micro- and macrovascular origin, J Vasc Res 44, 19-30, 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, (Bos taurus), calf	内皮 / Endothelial	Collagenase: 1000 u/ml； Collagenase Type 2: 1000 u/ml	PBS, CMF	Del Vecchio, P., Siflinger-Birnboim, A., Belloni, P., Holleran, L., Lum, H., and Malik, A.: Culture and Characterization of Pulmonary Microvascular Endothelial Cells, In Vitro Cell Dev Biol 28A, 711, 1992
牛	Calf	内皮 / Endothelial	Trypsin: 0.25%	HEPES	Vender, R.: Role of Endothelial Cells in the Proliferative Response of Cultured Pulmonary Vascular Smooth Muscle Cells to Reduced Oxygen Tension, In Vitro Cell Dev Biol 28A, 403, 1992
牛	Bovine, adult	脑动脉，内皮 / Cerebral artery，Endothelial	Collagenase: 0.2%	HBSS	Machi, T., Kassell, N.F., and Scheld, W.M.: Isolation and Characterization of Endothelial Cells From Bovine Cerebral Arteries, In Vitro Cell Dev Biol 26, 291, 1990
牛	Bovine	内皮 / Endothelial	Trypsin: 0.05%	Krebs Ringer solution	DeNucci, G., Gryglewski, R.J., Warner, T.D., and Vane, J.R.: Receptor-Mediated Release of Endothelium-Derived Relaxing Factor and Prostacyclin From Bovine Aortic Endothelial Cells Is Coupled, Proc Natl Acad Sci U S A 85, 2334, 1988
牛	Bovine	内皮，肺动脉 / Endothelial, pulmonary artery	Collagenase: 0.1%	CMF-Dulbecco's PBS	Martin, T.: Formation of Diacylglycerol by a Phospholipase D-phosphatidate Phosphatase Pathway Specific for Phosphatidylcholine in Endothelial Cells, Biochim Biophys Acta 962, 282, 1988
牛	Bovine	内皮 / Endothelial	Collagenase Type 2: 0.1%	PBS	Carson, M.P. and Haudenschild, C.C.: Microvascular Endothelium and Pericytes: High Yield, Low Passage Cultures, In Vitro Cell Dev Biol 22, 344, 1986
牛	Bovine	内皮，主动脉 / Endothelial，Aortic	Collagenase Type 2: 0.1%	DMEM	Kinsella, Michael G. and Wight, Thomas N.: Modulation of Sulfated Proteoglycan Synthesis by Bovine Aortic Endothelial Cells During Migration, J Cell Biol 102, 679, 1986
牛	Bovine	内皮 / Endothelial	Collagenase: 0.5%	DMEM/Ham's F-12	Gospodarowicz, D., Massoglia, S., Cheng, J., and Fujii, D.: Effect of Fibroblast Growth Factor and Lipoproteins on the Proliferation of Endothelial Cells Derived From Bovine Adrenal Cortex, Brain Cortex, and Corpus Luteum Capillaries, J Cell Physiol 127, 121, 1986
牛	Bovine	内皮，脑动脉 / Endothelial，Brain arteries	Collagenase Type 2: 0.2%	Dulbecco's PBS	Goetz, I.., Warren, J., Estrada, C., Roberts, E., and Krause, D.: Long-Term Cultivation of Arterial and Capillary Endothelium From Adult Bovine Brain, In Vitro Cell Dev Biol 21, 172, 1985
牛	Calf	内皮，平滑肌 / Endothelial，Smooth muscle	Collagenase: 0.75%	DMEM	Voyta, J., Via, D., Butterfield, C., and Zetter, B.: Identification and Isolation of Endothelial Cells Based on Their Increased Uptake of Acetylated-Low Density Lipoprotein, J Cell Biol 99, 2034, 1984
牛	Bovine	内皮，角膜 / Endothelial，Corneal	Trypsin: 0.2%	PBS: DMEM	Scott, D., Murray, J., and Barnes, M.: Investigation of the Attachment of Bovine Corneal Endothelial Cells, Exp Cell Res 144, 472, 1983
牛	Bovine, 2-3 weeks	内皮，肺动脉 / Endothelial，Pulmonary artery	Collagenase Type 1: 0.2%	RPMI 1640 w/1% Fetal Bovine Serum	Lee, S., Douglas, W., Deneke, S., and Fanburg, B.: Ultrastructural Changes in Bovine Pulmonary Artery Endothelial Cells Exposed to 80% O2 In Vitro, In Vitro 19, 714, 1983
牛	Bovine	内皮，角膜 / Endothelial, Corneal	Trypsin: 0.05%	0.01M Phosphate buffer with 0.02% EDTA；0.9% NaCl (See Reference)	Robinson, J. and Gospodarowicz, D.: Glycosaminoglycans Synthesized by Cultured Bovine Corneal Endothelial Cells, J Cell Physiol 117, 368, 1983
牛	Bovine	内皮，锁骨下静脉 / Endothelial，Subclavian vein	Collagenase Type 1: 0.10%	PBS	Olander, J., Marasa, J., Kimes, R., Johnston, G., and Feder, J.: An assay measuring the stimulation of several types of bovine endothelial cells by growth factor(s) derived from cultured human tumor cells, In Vitro 18, 99, 1982
牛	Bovine	主动脉，肺动脉 / Aortic，Pulmonary artery	Collagenase Type 2: 0.10%	PBS	Makarski, J. S.: Stimulation of cyclic AMP production by vasoactive agents in cultured bovine aortic and pulmonary artery endothelial cells, In Vitro 17, 450, 1981
牛	Calf, fetal, 4-9 months	内皮 / Endothelial	Collagenase Type 1: 0.25%	PBS	Rosen, E., Mueller, S., Noveral, J., and Levine, E.: Proliferative Characteristics of Clonal Endothelial Cell Strains, J Cell Physiol 107, 123, 1981
牛	Bovine, adult	主动脉 / Aorta	Collagenase Type 1: 125 u/ml	Dulbecco's PBS with calcium and magnesium	Cotta-Pereira, G., Sage, H., Bornstein, P., Ross, R., and Schwartz, S.: Studies of Morphologically Atypical ("Sprouting") Cultures of Bovine Aortic Endothelial Cells. Growth Characteristics and Connective Tissue Protein Synthesis, J Cell Physiol 102, 183, 1980
牛	Bovine, young	肺动脉 / Pulmonary artery	Collagenase: 0.1%	Medium 199	Ryan, U., Mortara, M., and Whitaker, C.: Methods for Microcarrier Culture of Bovine Pulmonary Artery Endothelial Cells Avoiding the Use of Enzymes, Tissue Cell 12, 619, 1980
牛	Bovine	包皮 / Foreskin	Collagenase: 0.5%	Dulbecco's MEM w/10% calf serum	Folkman, J., Haudenschild, C. C., and Zetter, B. R.: Long-term Culture of Capillary Endothlial Cells, Proc Natl Acad Sci U S A 76, 5217, 1979
牛	Bovine	大隐静脉，主动脉 / Saphenous Vein，Aorta	Collagenase: 0.01%	PBS	Eskin, S., Sybers, H., Trevino, L., Lie, J., and Chimoskey, J.: Comparison of Tissue-Cultured Bovine Endothelial Cells from Aorta and Saphenous Vein, In Vitro 14, 903, 1978
牛	Bovine	内皮，胸主动脉，隐静脉 / Endothelial，Thoracic aorta，Saphenous veins	Collagenase Type 2: 0.1%	PBS	Schwartz, S.M.: Selection and Characterization of Bovine Aortic Endothelial Cells, In Vitro 14, 966, 1978
牛	Bovine	肺动脉 / Pulmonary artery	Collagenase Type 2: 0.25%	Puck's solution	Ryan, U.S., Clements, E., Habliston, D., and Ryan, J.W.: Isolation And Culture of Pulmonary Artery Endothelial Cells, Tissue Cell 10, 535, 1978
牛	Calf	内皮 / Endothelial	Collagenase Type 2: 0.1%	DMEM	Howard, B., Macarak, E., Gunson, D., and Kefalides, N.: Characterization of the Collagen Synthesized by Endothelial Cells in Culture, Proc Natl Acad Sci U S A 73 (7), 2361, 1976
犬/狗	Dog	微血管 / Microvessels	Collagenase Type 4: 0.1%	DMEM	Gerhart, D. Z., Broderius, M. A., and Drewes, L. R.: Cutlured Human and Canine Endothelial Cells from Brain Microvessels, Brain Res Bull 21, 785, 1988
犬/狗	Dog, mongrel, adult	内皮，颈静脉 / Endothelium，Jugular vein	Trypsin: 0.1%	Earle's PBS, CMF	Ford, J., Burkel, W., and Kahn, R.: Isolation of Adult Canine Venous Endothelium for Tissue Culture, In Vitro 17, 44, 1981
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
人	Human	血管内皮 / Vascular endothelial	Collagenase Type 1: 0.1%	HBSS	Ganguly, A., Zhang, H., Sharma, R., Parsons, S. and Patel, K.: Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions., J Vis Exp 66, e4032, 2012
人	Human, 18-68 yr	角膜内皮 / Corneal endothelial	Collagenase: 0.2%； Neutral Protease: 1.0%	DMEM/F12	Li Wei, Sabater Alfonso L, Chen Ying-Ting, Hayashida Yasutaka, Chen Szu-Yu, He Hua, Tseng Scheffer C G: A novel method of isolation, preservation, and expansion of human corneal endothelial cells, Inv Ophthal Visual Sci 48, 614-20, 2007
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	血管内皮细胞 / HUVEC	Collagenase Type 1: 0.1%	HBSS	Davis, J., Crampton, S. and Hughes. C.: Isolation of Human Umbilical Vein Endothelial Cells (HUVEC)., J Vis Exp 3, 183, 2007
人	Human	血管内皮细胞 / HUVEC	Collagenase Type 4: 0.1%	RPMI 1640	Silva AP, Kaufmann JE, Vivancos C, Fakan S, Cavadas C, Shaw P, Brunner HR, Vischer U, and Grouzmann E.: Neuropeptide Y expression, localization and cellular transducing effects in HUVEC, Biol Cell 97(6), 457, 2005
人	Human	内皮 / Endothelial	Collagenase Type 2: 0.1%	DMEM	Patel, V., Logan, A., Watkinson, J., Uz-Zaman, S., Sheppard, M., Ramsden, J. and Eggo, M.: Isolation and Characterization of Human Thyroid Endothelial Cells., Am J Physiol Endocrinol Metab 284, E168, 2003
人	Human	食管微血管内皮 / Esophageal microvascular endothelial	Collagenase Type 2: 0.2%	MCDB-131	Rafiee, P., Ogawa, H., Heidemann, J., Li, M., Aslam, M., Lamirand, T., Fisher, P., Graewin, S., Dwinell, M., Johnson, C., Shaker, R. and Binion, D.: Isolation and Characterization of Human Esophageal Microvascular Endothelial Cells: Mechanisms of Inflammatory Activation., Am J Physiol Gastrointest Liver Physiol 285, G1277, 2003
人	Human	结肠内皮细胞 / Colonic endothelial cells	Collagenase Type 2: 0.25%	HBSS/5%FBS	Wang D., Lehman R., Donner D., Matli M., Warren R., and Welton M.: Expression and Endocytosis of VEGF and Its Receptors in Human Colonic Vascular Endothelial Cells, Am J Physiol/Gastro 282, G1088, 2002
人	Human	肺血管内皮细胞 / Pulmonary vascular endothelial cells	Neutral Protease: 1.18 u/ml； Elastase: 10 u/ml	M199	Muller AM, Hermanns MI, Skrzynski C, Nesslinger M, Muller KM, and Kirkpatrick CJ.: Expression of the endothelial markers PECAM-1, vWf, and CD34 in vivo and in vitro, Exp Mol Pathol 72, 221, 2002