worthington 內(nèi)皮細(xì)胞組織分離指南

發(fā)布者：艾美捷科技發(fā)布時(shí)間：2022-06-01

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原代細(xì)胞分離中，針對(duì)腦組織涉及到微血管內(nèi)皮、腦內(nèi)皮細(xì)胞、神經(jīng)元和小膠質(zhì)細(xì)胞、微血管等等的解離通常會(huì)使用酶，如何選擇合適的酶來(lái)完成對(duì)應(yīng)腦組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對(duì)腦相關(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(xiàn)，詳細(xì)匯總指南：

內(nèi)皮組織分離指南

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

內(nèi)皮細(xì)胞
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(xiàn)
牛	Bovine	牛臍帶 (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	肺動(dòng)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, (Bos taurus), calf	內(nèi)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	內(nèi)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	腦動(dòng)脈，內(nèi)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	內(nèi)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	內(nèi)皮，肺動(dòng)脈 / 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	內(nèi)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	內(nèi)皮，主動(dòng)脈 / 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	內(nèi)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	內(nèi)皮，腦動(dòng)脈 / 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	內(nèi)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	內(nèi)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	內(nèi)皮，肺動(dòng)脈 / 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	內(nèi)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	內(nèi)皮，鎖骨下靜脈 / 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	主動(dòng)脈，肺動(dòng)脈 / 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	內(nèi)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	主動(dòng)脈 / 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	肺動(dòng)脈 / 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	大隱靜脈，主動(dòng)脈 / 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	內(nèi)皮，胸主動(dòng)脈，隱靜脈 / 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	肺動(dòng)脈 / 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	內(nèi)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	內(nèi)皮，頸靜脈 / 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	冠狀動(dòng)脈內(nèi)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	血管內(nèi)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	角膜內(nèi)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	內(nèi)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	血管內(nèi)皮細(xì)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	血管內(nèi)皮細(xì)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	內(nèi)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	食管微血管內(nèi)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	結(jié)腸內(nèi)皮細(xì)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	肺血管內(nèi)皮細(xì)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