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Worthington組織消化指南
引言
組織消化酶主要用于組織培養(yǎng)和細(xì)胞生物學(xué)研究的原代細(xì)胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年，但是對(duì)其組織消化和細(xì)胞收集的作用機(jī)制卻沒有全面的理解。 由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機(jī)選擇或基于過去的經(jīng)驗(yàn)，而不是在*理解消化原理的基礎(chǔ)上選擇的消化方案，以及如何根據(jù)具體實(shí)驗(yàn)?zāi)康男拚桨敢垣@得更好的實(shí)驗(yàn)結(jié)果。
細(xì)胞分離程序的目的是zui大程度的獲得功能活性的單個(gè)細(xì)胞，而影響結(jié)果的原因有很多，包括（不限于）以下：
1、組織類型
2、物種來源
3、動(dòng)物年齡
4、遺傳改造情況（如knockouts）
5、所用的消化培養(yǎng)液
6、所用的酶
7、消化酶中的雜質(zhì)
8、所用消化酶的工作濃度
9、溫度
10、孵育時(shí)間
通常前四個(gè)原因沒辦法選擇，而剩下的原因一般由經(jīng)驗(yàn)決定以獲取zui適實(shí)驗(yàn)結(jié)果。
通過查閱文獻(xiàn)來選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實(shí)驗(yàn)數(shù)據(jù)搞得一頭霧水。這些巨大差異來源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性，以及過去常用的粗制、未提純的組織分離酶。另外，組織胞外基質(zhì)由各種蛋白、糖蛋白、脂類和糖脂組成，這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育（年齡）之間存在明顯的差異，這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶，如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。
本指南總結(jié)了目前常用組織消化酶的zui適消化和細(xì)胞收集知識(shí)和方法，描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟， 以便于制定一個(gè)*的細(xì)胞分離方案，同時(shí)列舉了一些相關(guān)的參考文獻(xiàn)。
下表列出多種組織的適宜消化酶（含參考文獻(xiàn)，以組織類型和物種分類，詳見鏈接）
脂肪組織 | 腎上腺組織 | 骨組織 | 腦組織 |
軟骨組織 | 結(jié)腸組織 | 內(nèi)皮組織 | 上皮組織 |
眼 | 心臟 | 腸 | 腎 |
肝 | 肺 | 淋巴結(jié) | 乳腺 |
其它組織 | 其他組織 | 肌組織 | 神經(jīng)組織 |
胰腺 | 腮腺 | 垂體 | 前列腺 |
生殖 | 動(dòng)物鱗片 | 皮膚 | 脾臟 |
干細(xì)胞 | 胸腺 | 甲狀腺/甲狀旁腺 | 扁桃體 |
腫瘤 |
|
|
|
注：使用Worthington公司消化酶的文獻(xiàn)不限于上表鏈接中所列，一般而言，Worthington公司的組織消化酶可用于大多組織類型的消化。
內(nèi)皮細(xì)胞組織消化手冊(cè)
| 物種 | 詳細(xì)來源 | 細(xì)胞名稱 | 消化用酶 | 培養(yǎng)基 | 參考文獻(xiàn) |
| 牛 | 牛 | 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 | 牛 | Pulmonary artery endothelial and smooth muscle cells | Collagenase: 0.04-0.05% | 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 |
| Soybean Trypsin Inhibitor: 0.04% | |||||
| 牛 | 牛, (歐洲牛), 牛犢 | Endothelial | Collagenase: 1000 u/ml | PBS, CMF | Del Vecchio, P.J., Siflinger-Birnboim, A., Belloni, P.N., Holleron, L.A., Lum, H., and Malik, A.B.: Culture and Characterization of Pulmonary Microvascular Endothelial Cell, In Vitro Cell Dev Biol 28A, 711, 1992 |
| Pulmonary | |||||
| 牛 | 牛犢 | 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 | 牛, 成年的 | Cerebral artery | 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 |
| Endothelial | |||||
| 牛 | 牛 | 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 |
| Aortic | |||||
| 牛 | 牛 | 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 | 牛 | 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 |
| Aorta | |||||
| 牛 | 牛 | Endothelial | 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 |
| Aortic | |||||
| 牛 | 牛 | 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 | 牛 | Endothelial | Collagenase Type 2: 0.2% | Dulbecco's PBS | Goetz, I.., Warren, J., Estrada, C., Roberts, E., and Krause, D.: Long-Term C*tion of Arterial and Capillary Endothelium From Adult Bovine Brain, In Vitro Cell Dev Biol 21, 172, 1985 |
| Brain arteries | |||||
| 牛 | 牛犢 | Endothelial | 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 |
| Smooth muscle | |||||
| 牛 | 牛 | Endothelial | 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 |
| Corneal | |||||
| 牛 | 牛, 2-3 周 | Endothelial | 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 |
| Pulmonary artery | |||||
| 牛 | 牛 | Endothelial, Corneal | Trypsin: 0.05% | 0.01M Phosphate buffer with 0.02% EDTA | Robinson, J. and Gospodarowicz, D.: Glycosaminoglycans Synthesized by Cultured Bovine Corneal Endothelial Cells, J Cell Physiol 117, 368, 1983 |
| 0.9% NaCl (See Reference) | |||||
| 牛 | 牛 | Endothelial | 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 |
| Subclavian vein | |||||
| 牛 | 牛 | Aortic | 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 |
| Pulmonary artery | |||||
| 牛 | 牛犢, 胎兒, 4-9 個(gè)月 | 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 | 牛, 成年的 | 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 | 牛, 年幼的 | 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 Cell12, 619, 1980 |
| 牛 | 牛 | 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 | 牛 | Saphenous Vein | 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 |
| Aorta | |||||
| 牛 | 牛 | Endothelial | Collagenase Type 2: 0.1% | PBS | Schwartz, S.M.: Selection and Characterization of Bovine Aortic Endothelial Cells,In Vitro 14, 966, 1978 |
| Thoracic aorta | |||||
| Saphenous veins | |||||
| 牛 | 牛 | 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 | 牛犢 | 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 | 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 |
| (包括人類) | |||||
| 犬 | 狗, 雜種狗, 成年的 | Endothelium | 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 |
| Jugular vein | |||||
| 豚鼠 | 豚鼠, 雌性, 300-350克 | 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 |
| 人類 | 人類 | Vascular endothelial | Collagenase Type 1: 0.1% | HBSS | Ganguly, A., Zhang, H., Sharma, R., Parsons, S. and Pa, 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 |
| 人類 | 人類, 18-68 yr | Corneal endothelial | Collagenase: 0.2% | 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 |
| Neutral Protease: 1.0% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | Endothelial | Collagenase Type 2: 0.1% | DMEM | Pa, 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 |
| 人類 | 人類 | 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 Physiol285, G1277, 2003 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | Pulmonary vascular endothelial cells | Neutral Protease: 1.18 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 |
| Elastase: 10 u/ml | |||||
| 人類 | 人類 | HUVEC | Collagenase Type 2: 0.1% | PBS | Takano Manabu, Meneshian Avedis, Sheikh Emran, Yamakawa Yasuhiko, Wilkins Kirsten Bass, Hopkins Elise A, Bulkley Gregory B: Rapid upregulation of endothelial P-selectin expression via reactive oxygen species generation, Am J Physiol Heart Circ Physiol 283, H2054-61, 2002 |
| 人類 | 人類 | Endothelial | Trypsin: 2% | PBS | Goolcharran, C., Cleland, J., Keck, R., Jones, A., and Borchardt, R.: Comparsion of the Rates of Deamidation, Diketopiperazine Formation and Oxidation in Recombinant Human Vascular Endothelial Growth Factor and Model Peptides,AAPS PharmSci 2 (1), 5, 2000 |
| 人類 | 人類 | HUVEC, porcine pulmonary arterial endothelial cells | Collagenase Type 2: 0.2% | DMEM | Kwak HJ, Lee SJ, Lee YH, Ryu CH, Koh KN, Choi HY, and Koh GY.: Angiopoietin-1 inhibits irradiation- and mannitol-induced apoptosis in endothelial cells, Circulation 101(19), 2317, 2000 |
| 人類 | 人類 | Vascular endothelial cells | Collagenase: 0.1% | DMEM | Schonbeck U, Sukhova GK, Graber P, Coulter S, Libby P: Augmented expression of cyclooxygenase-2 in human atherosclerotic lesions, Am J Pathol 155, 1281-91, 1999 |
| 人類 | 人類 | Hepatic endothelial cells | Collagenase: 0.2% | DMEM | Sanyal AJ, and Mirshahi F.: A simplified method for the isolation and culture of endothelial cells from pseudointima of transjugular intrahepatic portasystemic shunts, Lab Invest 78(11), 1469, 1998 |
| 人類 | 人類 | Foreskin microvascular endothelial | Trypsin: 0.3% | HBSS | Wojta J, Gallicchio M, Zoellner H, Filonzi E, Hamilton J, McGrath K: Interleukin-4 stimulates expression of urokinase-type-plasminogen activator in cultured human foreskin microvascular endothelial cells, Blood 81, 3285-92, 1993 |
| 人類 | 人類 | Endothelial | Trypsin: 0.3.% | HBSS | Lee, K, Lawley, T, Xu, Y, and Swerlick, R.: VCAM-1-, ELAM-1-, and ICAM-1-Independent Adhesion of Melanoma Cells to Cultured Human Dermal Microvascular Endothelial Cells, J Invest Dermatol 98, 79, 1992 |
| (see reference) | |||||
| 人類 | 人類 | Vascular endothelial | Neutral Protease: 0.15% | M-199 | Farber HW, Antonov AS, Romanov YA, Smirnov VN, Scarfo LM, Beer DJ: Cytokine secretion by human aortic endothelial cells is related to degree of atherosclerosis,Am J Physiol 262, H1088-95, 1992 |
| Trypsin: 0.25% | |||||
| 人類 | 人類 | Umbilical vein | Collagenase: 0.1% | Cord Buffer | Grant, D.S., Lelkes, P.I., Fukuda, K., Kleinman, H.K.: Intracellular Mechanisms Involved in Basement Membrane Induced Blood Vessel Differentiation In Vitro, In Vitro Cell Dev Biol 27, 327, 1991 |
| HUVEC | (See Reference) | ||||
| 人類 | 人類 | Crypt cells | Collagenase: 125 u/ml | RMPI 1640 | Whitehead, R., and Eeden, P.: A Method For the Prolonged Culture of Colonic Epithelial Cells, J Tiss Cul Meth 13, 103, 1991 |
| 人類 | 人類 | Endothelial | Collagenase Type 1: 0.2% | Medium 199 | Fischer, E., Stingl, A., and Kirkpatrick, C.: Migration Assay for Endothelial Cells in Multiwells Application to Studies on the Effect of Opioids, J Immunol Methods 128, 235, 1990 |
| 人類 | 人類 | Human umbilical vein endothelial cells | Collagenase Type 2: 75 u/ml | M199 | Muller WA, Ratti CM, McDonnell SL, Cohn ZA: A human endothelial cell-restricted, externally disposed plasmalemmal protein enriched in intercellular junctions, J Exp Med 170, 399-414, 1989 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | Endothelial/HUVEC | Trypsin: 0.3% | HBSS/PBS, Medium 199 | Kubota, Y., Kleinman, H., Martin, G., and Lawley, T.: Role of Laminin and Basement Membrane in Morphological Differentiation of Human Endothelial Cells into Capillary-like Structures, J Cell Biol 107, 1589, 1988 |
| Foreskin & umbilical cord | (see reference) | ||||
| 人類 | 人類 | Umbilical cord | Collagenase: 0.1% | HEPES | Hoshi, H., Kan, M., Chen, J., and McKeehan, W.: Comparative Endocrinology-Paracrinology-Autocrinology of Human Adult Large Vessel Endothelial and Smooth Muscle Cells, In Vitro Cell Dev Biol 24 (4), 309, 1988 |
| Smooth muscle | |||||
| 人類 | 人類 | Endothelial | Collagenase Type 2: 0.1% | PBS, CMF | Sharefkin, J.B., Fairchild, K.D., Albus, R.A., Cruess, D.F., and Rich, N.M.: The Cytoxic Effect of Surgical Glove Powder Particles on Adult Human Vascular Endothelial Cell Cultures: Implications for Clinical Uses of Tissue Culture Techniques, J Surg Res 41, 463, 1986 |
| Saphenous vein | |||||
| 人類 | 人類 | Endothelial | Collagenase: 0.1% | HEPES | Hoshi, H., and McKeehan, W.: Isolation, Growth Requirements, Cloning, Prostacylclin Production and Life-Span of Human Adult Endothelial Cells in Low Serum Culture Medium, In Vitro Cell Dev Biol 22 (1), 51, 1986 |
| 人類 | 人類 | Endothelial | Trypsin: 0.3% | PBS | Marks, R.M., Czerniecki, M., and Penny, R.: Human Dermal Microvascular Endothelial Cells: An Improved Method for Tissue Culture and Description of Some Singular Properties in Culture, In Vitro Cell Dev Biol 21, 627, 1985 |
| Dermal | |||||
| 人類 | 人類 | Fibroblasts | Hyaluronidase: 0.10% | DMEM | Gordon, P., Sussman, I., and Hatcher, V.: Long-Term Culture of Human Endothelial Cells, In Vitro 19, 661, 1983 |
| Foreskin | |||||
| 人類 | 人類 | Iliac arteries | Collagenase: 0.25% | PBS w/Ca++, Mg++, & BSA | Glassberg, M., Bern, M., Coughlin, S., Haudenschild, C., Hoyer, L., and Antoniades, H.: Cultured Endothelial Cells Derived from the Human Iliac Arteries,In Vitro 18, 859, 1982 |
| (see reference) | |||||
| 人類 | 人類, 成年的 | Peripheral blood mononuclear | Collagenase: 0.25% | RPMI 1640 | Ashida, E., Johnson, A., and Lipsky, P.: Human endothelial cell-lymphocyte interaction. Endothelial cells function as accessory cells necessary for mitogen-induced human T lymphocyte activation in vitro, J Clin Invest 67, 1490, 1981 |
| Monocytes | |||||
| T cells | |||||
| Endothelial | |||||
| 人類 | 人類 | Microvascular endothelial | Neutral Protease: at 1000 u/ml | Konigsberg's modification of HBSS | Sherer, G., Fitzharris, T., Faulk, W., and LeRoy, E.: C*tion of Microvascular Endothelial Cells from Human Preputial Skin, In Vitro 16, 675, 1980 |
| Neonatal foreskins | (See Reference) | ||||
| 人類 | 人類 (成年人和小孩) | 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 |
| 人類 | 人類 | Umbilical vein | Trypsin: 100 礸/ml | Tris-HCl,0.2 M | Jaffe, E.A., Minick, C.R., Adelman, B., Becker, C.G., and Nachman, R.: Synthesis of Basement Membrane Collagen By Cultured Human Endothelial Cells, J Exp Med 144, 209, 1976 |
| 人類 | 人類 | Umbilical vein | Collagenase Type 1: 125 u/ml | Dulbecco's PBS | Gimbrone Jr., M.A.: Culture of Vascular Endothelium, Prog Hemost Thromb 3, 1, 1976 |
| 人類 | 人類 | Umbilical cord | Collagenase: 0.1% | Dulbecco's PBS | Gimbrone, M.A., Cotran, R.S., and Folkman, J.: Human Vascular Endothelial Cells in Culture: Growth and DNA Synthesis, J Cell Biol 60, 673, 1974 |
| 人類 | 人類 | Umbilical cord | Collagenase: 0.2% | Cord buffer | Jaffe, E., Nachman, R., Becker, C., and Minick, C.: Culture of Human Endothelial Cells Derived from Umbilical Veins. Identification by Morphologic and Immunologic Criteria, J Clin Invest 52, 2745, 1973 |
| (See Reference) | |||||
| 人類 | 人類 | Umbilical cord | Trypsin NF 1:250: 0.125% | Saline, normal | Lewis, L.J., Haok, J.C., Maca, R.D., and Fry, G.L.: Replication of Human Endothelial Cells in Culture, Science 181, 452, 1973 |
| 人類 | 人類 | Umbilical cord | Trypsin NF 1:250: 0.25% | CMF solution | Fryer, D.G., Birnbaum, G., and Luttrell, C.N.: Human Endothelium in Cell Culture, J Atheroscler Res 6, 151, 1966 |
| 小鼠 | 小鼠 | Endothelial lung | Collagenase Type 1: 0.2% | DMEM | Wang, J., Sun, C., Gerdes, N., Liu, C., Liao, M., Liu, J., Shi, M., He, A., Zhou, Y., Sukhova, G., Chen, H., Cheng, X., Kuzuya, M., Murohara, T., Zhang, J., Cheng, X., Jiang, M., Shull, G. and Rogers, S.: Interleukin 18 Function in Atherosclerosis is Mediated by the Interleukin 18 Receptor and the Na-Cl Co-Transporter., Nat Med 21, 820-6, 2015 |
| 小鼠 | 小鼠 | Tumor endothelial | Collagenase Type 1: 0.2% | HBSS | Kazerounian, S., Gerald, D., Huang, M., Chin, R, Udayakumar, D, Zheng, N., O'Donnell, R., Perruzzi, C., Mangiante, L., Pourat, J., Phung, T., Bravo-Nuevo, A., Shechter, S., McNamara, S. and Duhadaway, K.: RhoB Differentially Controls Akt Function in Tumor Cells and Stromal Endothelial Cells During Breast Tumorigenesis., Cancer Res 73, 50, 2013 |
| 小鼠 | 小鼠, 8-14 week | Endothelial | Collagenase Type 4: 0.2% | HBSS | Imoukhuede, P. and Popel, A.: Expression of VEGF Receptors on Endothelial Cells in Mouse Skeletal Muscle., PLoS ONE 7, e44791, 2012 |
| 小鼠 | 小鼠, 7-10 周 | Liver endothelial | Collagenase: 0.03% | DMEM | Follenzi, A., Benten, D., Novikoff, P., Faulkner, L., Raut, S. and Gupta, S.:Transplanted Endothelial Cells Repopulate the Liver Endothelium and Correct the Phenotype of Hemophilia A Mice., J Clin Invest 118, 935, 2008 |
| 小鼠 | 小鼠, 4 周 | Endothelial kidney | Collagenase Type 1: 0.1% | DMEM | Kondo, S., Scheef, E., Sheibani, N. and Sorenson, C.: PECAM-1 Isoform-Specific Regulation of Kidney Endothelial Cell Migration and Capillary Morphogenesis., Am J Physiol Cell Physiol 292, C2070, 2007 |
| 小鼠 | 小鼠, 胚胎或卵黃囊 | Endothelial | Collagenase Type 3: 200 u/ml | PBS | Braren Rickmer, Hu Huiqing, Kim YungHae, Beggs HilaryE, Reichardt LouisF, Wang Rong: Endothelial FAK is essential for vascular network stability, cell survival, and lamellipodial formation, J Cell Biol 172, 151-62, 2006 |
| Deoxyribonuclease I: 0.001% | |||||
| 小鼠 | 小鼠, 初生的 | Microvascular endothelial | Neutral Protease: 0.005% | DMEM | Cha, S., Talavera, D., Demir, E., Nath, A. and Sierra-Honigmann, M.: A Method of Isolation and Culture of Microvascular Endothelial Cells from Mouse Skin.,Microvasc Res 70, 198, 2005 |
| Collagenase Type 1: 4% | |||||
| 小鼠 | 小鼠, 4 周 | Retinal endothelial cells | Collagenase Type 1: 0.1% | DMEM | Su X, Sorenson CM, and Sheibani N.: Isolation and characterization of murine retinal endothelial cells, Mol Vis 9, 171, 2003 |
| 小鼠 | 小鼠, 雄性 | Endothelial cells from lymph node | Collagenase Type 1: 0.1% | PBS | Izawa D, Tanaka T, Saito K, Ogihara H, Usui T, Kawamoto S, Matsubara K, Okubo K, and Miyasaka M: Expression profile of active genes in mouse lymph node high endothelial cells, Int Immunol 11(12), 1989, 1999 |
| 小鼠 | 小鼠, 2-4 周 | Cerebrovascular Endothelial | Collagenase/Dispase: 0.1% | PBS | Sapatino, B., Welsh, C., Smith, C., Bebo, B., and Linticum, D.: Cloned Mouse Cerebrovascular Endothelial Cells That Maintain Their Differentiation Markers for Factor VIII, Low Density Lipoprotein, and Angiotensin-Converting Enzyme, In Vitro Cell Dev Biol 29A, 923, 1993 |
| 豬 | 豬, 6 個(gè)月 | Valvular endothelial | Neutral Protease: 2 u/ml | PBS | Balaoing, L., Post, A., Lin, A., Tseng, H., Moake, J. and Grande-Allen, K.: Laminin Peptide-Immobilized Hydrogels Modulate Valve Endothelial Cell Hemostatic Regulation., PLoS ONE 10, e0130749, 2015 |
| Collagenase Type 2: 60 u/ml | |||||
| 豬 | 豬 | Valvular endothelial | Collagenase Type 2: 300-600 u/ml | DMEM | Gould, R. and Butcher, J.: Isolation of Valvular Endothelial Cells., J Vis Exp 46, e2158, 2010 |
| 豬 | 豬 | Endothelial | Collagenase Type 1: | DMEM w/ 10% calf serum | Nugent, H., and Edelman, E.: Endothelial Implants Provide Long-Term Control of Vascular Repair in a Porcine Model of Arterial Injury, J Surg Res 99, 228, 2001 |
| (包括 牛) | |||||
| 豬 | 豬 | HUVEC, porcine pulmonary arterial endothelial cells | Collagenase Type 2: 0.2% | DMEM | Kwak HJ, Lee SJ, Lee YH, Ryu CH, Koh KN, Choi HY, and Koh GY.: Angiopoietin-1 inhibits irradiation- and mannitol-induced apoptosis in endothelial cells, Circulation 101(19), 2317, 2000 |
| 豬 | 豬, 6-7 個(gè)月 | Porcine pulmonary endothelial | Collagenase Type 1: 0.3% | RPMI 1640 | Hill-Kapturczak N, Kapturczak MH, Block ER, Pa JM, Malinski T, Madsen KM, and Tisher CC.: Angiotensin II-stimulated nitric oxide release from porcine pulmonary endothelium is mediated by angiotensin IV, J Am Soc Nephrol 10(3), 481, 1999 |
| 豬 | 豬 | Endothelial | Trypsin: 0.25% | Medium 199 | Shasby, S.: Endothelial Cells Grown On Permeable Membrane Supports, J Tiss Cul Meth 14, 247, 1992 |
| 豬 | 豬 | Endothelial | Trypsin: 0.05% | DMEM | Vischer, P., and Buddecke, E.: Alteration of Glycosytransferase Activities during Proliferation of C*ted Arterial Endothelial Cells and Smooth Muscle Cells, Exp Cell Res 158, 15, 1985 |
| 豬 | 豬 | Endothelial | Collagenase Type 2: 0.1% | Dulbecco-Vogt MEM w/o serum | Dickinson, E. and Slakey, L.: Plasma-derived Serum as a Selective Agent to Obtain Endothelial Cultures from Swine Aorta, In Vitro 18, 63, 1982 |
| Aortic | |||||
| 豬 | 豬, 30-40 公斤 | Endothelia | Collagenase Type 4: 0.025% | Medium 199 | Merrilees, M.J., and Scott, L.: Interaction of Aortic Endothelial and Smooth Muscle Cells in Culture, Atherosclerosis 39, 147, 1981 |
| Aortic | |||||
| 豬 | 豬, 20-30 周 | Endothelial | Collagenase: 0.1% | Medium 199 w/BSS and HEPES or NaHCO3 | Slater, D.N., and Sloan, J.M.: The Porcine Endothelial Cell in Tissue Culture,Atherosclerosis 21, 259, 1975 |
| Aortas | |||||
| Veins | |||||
| 豬 | 豬, 60-100 天 | Aorta | Trypsin: 0.1% | Phosphate buffer | Coulson, W.F.: The Effect Of Proteolytic Enzymes on the Tensile Strength of Whole Aorta and Isolated Aortic Elastin, Biochim Biophys Acta 237, 378, 1971 |
| (see reference) | |||||
| 兔 | 兔 | Corneal endothelial cells (CEC) | Hyaluronidase: 0.05% | DMEM | Choi, J., Ko, M., and Kay, E.: Subcellular Localization of the Expressed 18 kDa FGF-2 Isoform in Corneal Endothelial Cells, Mol Vis 6, 222, 2000 |
| 兔 | 兔, 2-3公斤 | Endothelial, aortic | Elastase: 0.2% | Hanks solution | Haley, N., Shio, H., Fowler, S.: Characterization of lipid-laden aortic cells from cholesterol-fed rabbits. I. Resolution of aortic cell populations by metrizamide density gradient centrifugation, Lab Invest 37, 287, 1977 |
| 大鼠 | 大鼠, SD, 雄性, 250-300 克 | Smooth muscle, aorta | Soybean Trypsin Inhibitor: 0.25% | HBSS with 0.2 mM Ca++ | Schwertschlag, U.S., and Whorton, A.R.: Plaet-Activating Factor-Induced Homologous and Heterologous Desensitization In Cultured Vascular Smooth Muscle Cells, J Biol Chem 263, 13791, 1988 |
| 大鼠 | 大鼠, SD, 雄性, 350 - 450 g | Lipocytes | Collagenase: 0.015% | DMEM/Ham's F-12 | Friedman, S. and Roll, F.: Isolation and Culture of Hepatic Lipocytes,Kupffer Cells, and Sinusoidal Endothelial Cells by Density Gradient Centrifugation with Stractan,Anal Biochem 161, 207, 1987 |
| Kupffer | |||||
| Sinusoidal endothelial | |||||
| 大鼠 | wistar大鼠, 雄性, 3 個(gè)月 | Endothelial | Pronase: 0.25% | HBSS | Nagelkenke, J., Barto, K., and VanBerkel, T.: In vivo and in vitro uptake and degradation of acetylated low density lipoprotein by rat liver endothelial, Kupffer, and parenchymal cells, J Biol Chem 258, 12221, 1983 |
| Kupffer | |||||
| Parenchymal | |||||
| 大鼠 | 京都大鼠, 雄性, 100 - 200 克 | Endothelial | Collagenase Type 2: 0.05% | HBSS | Diglio, C.A., Grammas, P., Filiberto Giacomelli, M.S., and Wiener, J.: Primary Culture of Rat Cerebral Microvascular Endothelial Cells, Lab Invest 46, 554, 1982 |
| Cerebral | |||||
| 大鼠 | 大鼠, 300 克, 和 豬, 30-40 公斤 | Endothelial | Trypsin: 0.05% | Medium 199 and 0.01M EDTA | Merrilees, M.J., and Scott, L.: Interaction of aortic endothelial and smooth muscle cells in culture. Effect on glycosaminoglycan levels, Atherosclerosis 39, 147, 1981 |
| Thoracic aorta | |||||
| 大鼠 | 大鼠 | 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 |
| 大鼠 | wistar大鼠, 成年的, 170 克 | Endothelial | Trypsin: 0.5% | BSS | Phillips, P., Kumar, P., Kumar, S., and Waghe, M.: Isolation And Characterization of Endothelial Cells From Rat And Cow Brain White Matter, J Anat 129, 261, 1979 |
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