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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和細胞生物學研究的原代細胞的分離和收集。盡管這些消化酶已經普遍應用多年，但是對其組織消化和細胞收集的作用機制卻沒有全面的理解。 由此導致的結果便是選擇一種消化方法大多是隨機選擇或基于過去的經驗，而不是在*理解消化原理的基礎上選擇的消化方案，以及如何根據具體實驗目的修正消化方案以獲得更好的實驗結果。
細胞分離程序的目的是zui大程度的獲得功能活性的單個細胞，而影響結果的原因有很多，包括（不限于）以下：
1、組織類型
2、物種來源
3、動物年齡
4、遺傳改造情況（如knockouts）
5、所用的消化培養液
6、所用的酶
7、消化酶中的雜質
8、所用消化酶的工作濃度
9、溫度
10、孵育時間
通常前四個原因沒辦法選擇，而剩下的原因一般由經驗決定以獲取zui適實驗結果。
通過查閱文獻來選擇*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驗數據搞得一頭霧水。這些巨大差異來源于組織胞外基質的復雜而動態的特性，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。
本指南總結了目前常用組織消化酶的zui適消化和細胞收集知識和方法，描述了標準的實驗步驟， 以便于制定一個*的細胞分離方案，同時列舉了一些相關的參考文獻。
下表列出多種組織的適宜消化酶（含參考文獻，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 | 結腸組織 | 內皮組織 | 上皮組織 |
眼 | 心臟 | 腸 | 腎 |
肝 | 肺 | 淋巴結 | 乳腺 |
其它組織 | 其他組織 | 肌組織 | 神經組織 |
胰腺 | 腮腺 | 垂體 | 前列腺 |
生殖 | 動物鱗片 | 皮膚 | 脾臟 |
干細胞 | 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。
內皮細胞組織消化手冊
| 物種 | 詳細來源 | 細胞名稱 | 消化用酶 | 培養基 | 參考文獻 |
| 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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