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世界*實驗材料供應商 Worthington Biochemical 正式授權上海起發為其中國代理， Worthington Biochemical 在一直是行業的*,一直為廣大科研客戶提供zui為的產品和服務,上海起發一直秉承為中國科研客戶帶來的產品，的服務，簽約 Worthington Biochemical 就是為了給廣大科研客戶帶來更加完善的產品和服務,您的滿意將是我們zui大的收獲
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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。
腎組織消化手冊
| 物種 | 詳細來源 | 細胞名稱 | 消化用酶 | 培養基 | 參考文獻 |
| 鳥類 | 鳥類, 麻雀 | Tubule segments | Collagenase Type 2: 0.1% | HBSS | Goldstein D., Reddy V., and Plaga K.: Second Messenger Production in Avian Medullary Nephron Segments in Response to Peptide Hormones, Am J Physiol 276, R847, 1999 |
| 鳥類 | 鳥類, 雞 | Tubule and glomeruli | Collagenase Type 1: 0.1% | RPMI 1640 | Reich, C. and Bonar, R.: Separation of Avian Kidney Tubules and Glomeruli for In Vitro Culture, Meth Cell Sci 7, 97, 1982 |
| 牛 | 牛 | Papillary duct | Hyaluronidase: 0.2% | Keri's buffer | Husted, R., Hayashi, M., and Stokes, J.: Characteristics of Papillary Collecting Duct Cells in Primary Culture, Am J Physiol 255, F1160, 1988 |
| HEPES buffered saline | |||||
| 犬 | 犬 | Renal adipose derived cells | Collagenase Type 1: 0.3% | DMEM | Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011 |
| 犬 | 狗, 1-5 個月 | Proximal tubules | Collagenase Type 1: 0.15% | Krebs Ringer bicarbonate buffer | States, B., Reynolds, R., Lee, J., and Segal, S.: Cystine Uptake By Cultured Cells Originating From Dog Proximal Tubule Segments, In Vitro Cell Dev Biol 26, 105, 1990 |
| 犬 | 狗, 雜種狗, 20 公斤 | Proximal tubular | Deoxyribonuclease I: 0.0125% | (see reference) | Yau, C., Rao, L., and Silverman, M.: Sugar Uptake Into a Primary Culture of Dog Kidney Proximal Tubular Cells, Can J Physiol Pharmacol 63, 417, 1985 |
| 魚 | 美洲擬鰈 (鈍吻黃蓋鰈) | Renal tubule | Trypsin: 0.2% | CMF solution | Dickman, K. and Renfro, J.: Primary Culture of Flounder Renal Tubule Cells:Transepithelial Transport, Am J Physiol 251, 424, 1986 |
| 200 - 500 克 | (see reference) | ||||
| 豚鼠 | 豚鼠, 雄性, 100 克 | Single cells | Trypsin: 0.25% | CF salt solution | Phillips, H.: Dissociation of Single Cells from Lung or Kidney Tissue with Elastase, In Vitro 8, 101, 1972 |
| 倉鼠 | 倉鼠 | Papillary duct | Hyaluronidase: 0.2% | Keri's buffer | Husted, R., Hayashi, M., and Stokes, J.: Characteristics of Papillary Collecting Duct Cells in Primary Culture, Am J Physiol 255, F1160, 1988 |
| (包括SD大鼠, Wistar大鼠, 150-225 克, 兔, 牛) | HEPES buffered saline | ||||
| 人類 | 人類 | Renal adipose derived cells | Collagenase Type 1: 0.3% | DMEM | Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011 |
| 人類 | 人類, 成年人 | Renal tumor cells and proximal tubular epithelial | Collagenase Type 2: 0.1% | DMEM/F12 | Valente, M., Henrique, R., Costa, V., Jeronimo, C., Carvalho, F., Bastos, M., Guedes de Pinho, P. and Carvalho, M.: A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens., PLoS ONE 6, e19337, 2011 |
| 人類 | 人類 | Renal cells | Collagenase Type 4: 300 u/ml | HBSS | Presnell, S., Bruce, A., Wallace, S., Choudhury, S., Genheimer, C., Cox, B., Guthrie, K., Werdin, E., Tatsumi-Ficht, P., Ilagan, R., Kelley, R., Rivera, E., Ludlow, J., Wagner, B., Jayo, M. and Bertram, T.: Isolation, Characterization, and Expansion Methods for Defined Primary Renal Cell Populations from Rodent, Canine, and Human Normal and Diseased Kidneys., Tiss Eng 17, 261, 2011 |
| Neutral Protease: see reference | |||||
| 人類 | 人類, 成年人 | Renal proximal tubule and cortical fibroblasts | Collagenase Type 2: 383 u/ml | DMEM/F-12 | Johnson, D., Saunders, H., Johnson, F., Huq, S., Field, M., and Pollock, C.: Cyclosporin exerts a direct fibrogenic effect on human tubulointerstitial cells: roles of insulin-like growth factor I, transforming growth factor beta1, and plaet-derived growth factor, J Pharmacol Exp Ther 289, 535-42, 1999 |
| 人類 | 人類 | Renal cortex | Trypsin: 0.1% | Tissue Culture Grade Water | McAteer, J, Kempson, S., and Evan, A: Culture of Human Renal Cortex Epithelial Cells, J Tiss Cul Meth 13, 143, 1991 |
| 人類 | 人類 | Papillary duct | Collagenase: 400 u/ml | Eagle's MEM-HEPES buffer w/L-glutamine | Trifillis, A. and Kahng, M.: Characterization of an In Vitro System of Human Renal Papillary Collecting Duct Cells, In Vitro Cell Dev Biol 26, 441, 1990 |
| 人類 | 人類 | Mesangial | Trypsin: 0.25% | DMEM/Ham's F-12 | Heieren, M., van der Woude, F., and Balfour Jr., H.: Cytomegalovirus Replicates Efficiently in Human Kidney Mesangial Cells, Proc Natl Acad Sci U S A 85, 1642, 1988 |
| 人類 | 人類, 成年人 | Tubular | Collagenase: 250 u/ml | PBS | Yang, A., Gould-Kostka, J., and Oberley, T.: In Vitro Growth and Differentiation of Human Kidney Tubular Cells on a Basement Membrane Substrate, In Vitro Cell Dev Biol 23 (1), 34, 1987 |
| 人類 | 人類, 成年人, 14-66 歲 | Tubular | Collagenase: 100 u/ml | Joklik's MEM | Trifillis, A., Regec, A., and Trump, B.: Isolation, Culture, and Charaterization of Human Renal Tubular Cells, J Urol 133, 324, 1985 |
| 人類 | 人類 | Malignant | Papain: 0.009% | Sacks solution | Hemstreet, G., Enoch, P., and Pretlow, T.: Tissue Disaggregation of Human Renal Cell Carcinoma with Further Isopyknic and Isokinetic Gradient Purification, Cancer Res 40, 1043, 1980 |
| Stromal | |||||
| 人類 | 人類, 24 初生和 夭折的 | Renal | Trypsin: 0.25% | See reference | De Oca, H., and Malinin, T.: Dispersion and C*tion of Renal Cells After Short-Term Storage of Kidneys, J Clin Microbiol 2, 243, 1975 |
| (包括兔) | |||||
| 猴 | 猴 | Kidney | Trypsin: 0.01% - 0.00001% | Eagle's MEM | Melnick, J., and Wallis, C.: Problems Related to the Use of Serum and Trypsin in the Growth of Monkey Kidney Cells, Dev Biol 37, 77, 1976 |
| 小鼠 | 小鼠 | Collecting duct epithelial | Collagenase Type 2: 0.1% | DMEM | DiRocco, D., Kobayashi, A., Taketo, M., McMahon, A. and Humphreys, B.: Wnt4/尾-catenin Signaling in Medullary Kidney Myofibroblasts., J Am Soc Nephrol 24, 1399, 2013 |
| 小鼠 | 小鼠 | Glomeruli | Collagenase Type 4: 0.1% | RPMI 1640 | Kabgani, N., Grigoleit, T., Schulte, K., Sechi, A., Sauer-Lehnen, S., Tag, C., Boor, P., Kuppe, C., Warsow, G., Schordan, S., Mostertz, J., Chilukoti, R., Homuth, G., Endlich, N., Tacke, F. and Weiskirchen, R.: Primary Cultures of Glomerular Parietal Epithelial Cells or Podocytes with Proven Origin., PLoS ONE 7, e34907, 2012 |
| 小鼠 | 小鼠, 10-14 week | Proximal tubule | Collagenase Type 1: 0.1% | See reference | Park, J., Pasupulati, R., Feldkamp, T., Roeser, N. and Weinberg, J.: Cyclophilin D and the Mitochondrial Permeability Transition in Kidney Proximal Tubules after Hypoxic and Ischemic Injury., Am J Physiol/Renal 301, F134, 2011 |
| 小鼠 | 小鼠, 胚胎 | Kidney | Collagenase Type 1: 0.25% | HBSS | Brown, A., Blank, U., Adams, D., Karolak, M., Fetting, J., Hill, B. and Oxburgh, L.:Isolation and Culture of Cells from the Nephrogenic Zone of the Embryonic Mouse Kidney., J Vis Exp 50, e2555, 2011 |
| Pancreatin: 1.0% | |||||
| Deoxyribonuclease I: 1 u/ml | |||||
| 小鼠 | 小鼠,成年的 | Kidney Sphere-Derived | Collagenase Type 4: 0.1% | Iscove's Modified Dulbecco's | Huang, Y., Johnston, P., Zhang, B., Zakari, A., Chowdhry, T., Smith, R., Marban, E., Rabb, H. and Womer, K.: Kidney-Derived Stromal Cells Modulate Dendritic and T Cell Responses., J Am Soc Nephrol 20, 831-41, 2009 |
| 小鼠 | 小鼠, 雄性, 8-16 周 | Renal tubular epithelial | Collagenase: 200 u/ml | HBSS | Breggia, A. and Himmelfarb, J.: Primary Mouse Renal Tubular Epithelial Cells have Variable Injury Tolerance to Ischemic and Chemical Mediators of Oxidative Stress., Oxid Med Cell Longev 1, 33, 2008 |
| Soybean Trypsin Inhibitor: see reference | |||||
| 小鼠 | 小鼠 | Proximal tubule | Collagenase Type 2: 0.013% | see reference | Wright, J., Morales, M., Sousa-Menzes, J., Ornellas, D., Sipes, J., Cui, Y., Cui, I., Hulamm, P., Cebotaru, V., Cebotaru, L., Guggino, W. and Guggino, S.: Transcriptional Adaptation to Clcn5 Knockout in Proximal Tubules of Mouse Kidney., Physiol Genomics 33, 341, 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, 成年的 | Cortex, proximal tubule | Collagenase: 0.15% | DMEM | Syal Ashu, Schiavi Susan, Chakravarty Sumana, Dwarakanath Vangipuram, Quigley Raymond, Baum Michel: Fibroblast growth factor-23 increases mouse PGE2 production in vivo and in vitro, Am J Physiol/Renal 290, F450-5, 2006 |
| 小鼠 | 小鼠 | Kidney Mesenchymal | Collagenase: 250 u/ml | DMEM/F12 | Plotkin, M. and Goligorsky, M.: Mesenchymal Cells from Adult Kidney Support Angiogenesis and Differentiate into Multiple Interstitial Cell Types including Erythropoietin-Producing Fibroblasts., Am J Physiol Renal Physiol 291, F902-12, 2006 |
| Neutral Protease: 33.3 u/ml | |||||
| 小鼠 | 小鼠, 10-12 周 | Kidney | Collagenase Type 4: 0.075% | DMEM | Dekel, B., Zangi, L., Shezen, E., Reich-Zeliger, S., Eventov-Friedman, S., Katchman, H., Jacob-Hirsch, J., Amariglio, N., Rechavi, G., Margalit, R. and Reisner, Y.: Isolation and Characterization of Nontubular sca-1+lin- Multipotent Stem/Progenitor Cells from Adult Mouse Kidney., J Am Soc Nephrol 17, 3300, 2006 |
| 小鼠 | 小鼠, 雄性, 12-16 周 | Renal proximal tubule cells | Collagenase Type 2: 0.1% | DMEM/F-12 | Cunningham, R., Xiaofei, E., Steplock, D., Shenolikar, S. and Weinman, E.: Defective PTH Regulation of Sodium-Dependent Phosphate Transport in NHERF-1-/- Renal Proximal Tubule Cells and Wild-Type Cells Adapted to Low-Phosphate Media., Am J Physiol Renal Physiol 289, F933, 2005 |
| Soybean Trypsin Inhibitor: 0.25% | |||||
| 小鼠 | 小鼠, 雄性, 2 個月 | Cortical collecting duct | Collagenase: 54-178 u/ml | MEM | Sindic, A., Velic, A., Basoglu, C., Hirsch, J., Edemir, B., Kuhn, M. and Schlatter, E.:Uroguanylin and Guanylin Regulate Transport of Mouse Cortical Collecting Duct Independent of Guanylate Cyclase C., Kidney Int 68, 1008, 2005 |
| Protease: see reference | |||||
| 小鼠 | 小鼠, 6 周 | Glomerular mesangial cells | Collagenase Type 4: 0.1% | RPMI 1640 | Radeke HH, Janssen-Graalfs I, Sowa EN, Chouchakova N, Skokowa J, Loscher F, Schmidt RE, Heeringa P, Gessner JE.: Opposite regulation of type II and III receptors for immunoglobulin G in mouse glomerular mesangial cells and in the induction of anti-glomerular basement membrane (GBM) nephritis, J Biol Chem 277(30), 27535, 2002 |
| 小鼠 | 小鼠, 3-7 個月 | Fibroblasts, mesangial, smooth muscle | Trypsin: 0.25% | DMEM | Bradshaw AD, Francki A, Motamed K, Howe C, Sage EH: Primary mesenchymal cells isolated from SPARC-null mice exhibit altered morphology and rates of proliferation, Mol Biol Cell 10, 1569-79, 1999 |
| Collagenase: see reference | |||||
| Soybean Trypsin Inhibitor: .05% | |||||
| 小鼠 | 小鼠 | Epithelial | Collagenase Type 4: 1% | DMEM /F-12 | Taub, M: Renal Tubule Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 2,Doyle, A., Griffiths, J., and Newell, D., John Wiley & Sons, Ltd., 14B:6.1, 1995 |
| 小鼠 | 小鼠,單純的 SJL (H-2S) | Proximal tubular epithelial | Deoxyribonuclease I: 15 礸/ml | RPMI 1640 | Haverty, T., Kelly, C., Hines, W., Amenta, P., Watanabe, M., Harper, R., Kefalides, N., and Neilson, E.: Characterization of a Renal Tubular Epithelial Cell Line Which Secretes the Autologous Target Antigen of Autoimmune Experimental Interstitial Nephritis, J Cell Biol 107, 1359, 1988 |
| 豬 | 豬, 雜種 | Kidney | Collagenase Type 1: 0.1% | HBSS | Heussner, A. and Dietrich, D.: Primary Porcine Proximal Tubular Cells as an Alternative to Human Primary Renal Cells in Vitro: an Initial Characterization., BMC Cell Biol 14, 55, 2013 |
| 兔 | 兔 | Renal proximal tubular | Collagenase Type 1: 60 u/ml | DMEM/F12 | Nowak, G. and Bakajsova, D.: Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes., J Vis Exp 71, e4301, 2013 |
| 兔 | 兔, 新西蘭 | Proximal tubule cells | Collagenase Type 4: 0.005% | DMEM/F-12 | Taub, M.: Primary Kidney Proximal Tubule Cells., Methods Mol Biol 290, 231, 2005 |
| Soybean Trypsin Inhibitor: 0.00025% | |||||
| 兔 | 兔, 新西蘭 白, 雌性 | Nephron segments | Collagenase Type 2: 0.05% | Eagle's MEM | Schafer, J., Watkins, M., Li, L., Herter, P., Haxelmans, S., and Schlatter, E.: A Simplified Method for Isolation of Large Numbers of Defined Nephron Segments, Am J Physiol 273, F650, 1997 |
| Deoxyribonuclease I: 50 u/ml | |||||
| 兔 | 兔, 新西蘭 白, 雄性, 1.5 - 2.0 公斤 | Duct | Soybean Trypsin Inhibitor: 0.025% | Hank's Solution with calcium and HEPES | Naray-Fejes-Toth, A., and Fejes-Toth, G.: Immunoelection and Culture of Cortical Collecting Duct Cells, J Tiss Cul Meth 13, 179, 1991 |
| 兔 | 兔, 新西蘭 掰,雌性, 2-3 公斤 | Renal proximal tubules | Deoxyribonuclease I: 70 u/ml | Modified DME-F12 | Rodeheaver, D., Aleo, M., and Schnellmann, R.: Differences in Enzymatic and Mechanical Isolated Rabbit Renal Proximal Tubules: Comparison in Long-Term Incubation, In Vitro Cell Dev Biol 26, 898, 1990 |
| 兔 | 兔 | Papillary duct | Hyaluronidase: 0.2% | Keri's buffer | Husted, R., Hayashi, M., and Stokes, J.: Characteristics of Papillary Collecting Duct Cells in Primary Culture, Am J Physiol 255, F1160, 1988 |
| (包括, SD 大鼠和Wistar大鼠, 150-225克, 馬, 牛) | HEPES buffered saline | ||||
| 兔 | 兔, 新西蘭, 白色, 2-3 公斤 | MTALH cells | Collagenase: 0.1% | DMEM | Allen, M., Nakao, A., Sonnenburg, W., Burnatowska-Hledin, M., Speilman, W., and Smith, W.: Immunodissection of Cortical and Medullary Thick Ascending Limb Cells From Rabbit Kidney, Am J Physiol 255, F704, 1988 |
| RCCT cells | |||||
| 兔 | 兔, 新西蘭, 白色 | Collecting tubule | Trypsin: 0.05% | Kreb's Ringer buffer | Grenier, F., and Smith, W.: Formation of 6-keto-PGF-1aby Collecting Tubule Cells Isolated from Rabbit Renal Papillae, Prostaglandins 16, 759, 1978 |
| 兔 | 兔, 新西蘭, 白色, 雌性, 2.5 公斤 | Papillary collecting duct | Collagenase: 0.2% | (see reference) | Dworzack, D., and Grantham, J.: Preparation of Renal Papillary Collecting Duct Cells for Study, Nat Neurosci 8, 191, 1975 |
| 大鼠 | SD大鼠, 胎兒 | Fetal kidney | Collagenase Type 4: 0.1% | MEM | Gupta, A., Jadhav, S., Tripathy, N. and Nityanand, S.: Fetal Kidney Cells Can Ameliorate Ischemic Acute Renal Failure in Rats through Their Anti-Inflammatory, Anti-Apoptotic and Anti-Oxidative Effects., PLoS ONE 10, e0131057, 2015 |
| 大鼠 | Lewis大鼠, 雄性 | Renal adipose derived cells | Collagenase Type 1: 0.3% | DMEM | Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011 |
| 大鼠 | 大鼠 | Proximal tubule | Collagenase Type 2: 0.1% | see reference | Panico, C., Luo, Z., Damiano, S., Artigiano, F., Gill, P. and Welch, W.: Renal Proximal Tubular Reabsorption is Reduced in Adult Spontaneously Hypertensive Rats: Roles of Superoxide and Na+/H+ Exchanger 3., Hypertension 54, 1291, 2009 |
| 大鼠 | SD大鼠, 雄性, 200-250克 | Medullary thick ascending limb | Collagenase: 0.1% | HEPES-saline | Silva, G., Beierwaltes, W., and Garvin, J.: Extracellular ATP stimulates NO production in rat thick ascending limb, Hypertension 47, 563-7, 2006 |
| 大鼠 | Wistar大鼠, 300克 | Renal proximal tubules | Collagenase Type 2: 0.2% | DMEM/F12 | Deng Aihua, Miracle Cynthia M, Lortie Mark, Satriano Joseph, Gabbai Francis B, Munger Karen A, Thomson Scott C, Blantz Roland C: Kidney oxygen consumption, carbonic anhydrase, and proton secretion, Am J Physiol/Renal 290, F1009-15, 2006 |
| 大鼠 | 大鼠, Wistar, 雄性, 300-350 克 | Renal proximal tubules | Collagenase Type 2: 0.2% | DMEM/F-12 | Deng, A., Miracle, C., Suarez, J., Lortie, M., Satriano, J., Thomson, S., Munger, K. and Blantz, R.: Oxygen Consumption in the Kidney: Effects of Nitric Oxide Synthase Isoforms and Angiotensin II., Kidney Int 68, 723, 2005 |
| 大鼠 | 大鼠 | Cortical tubule cells | Collagenase: 0.6% | DMEM | Arystarkhova, E., Wetzel R., and Sweadner K.: Distribution and Oligomeric Association of Splice Forms of Na(+)-K(+)-ATPase Regulatory Gamma-subunit in Rat Kidney, Am J Physiol/Renal 282(3), F393, 2002 |
| 大鼠 | SD大鼠, 雄性 275-300 克 | Glomerular mesangial cells | Collagenase Type 1: 250 u/ml | DMEM | Amiri Farhad, Shaw Sean, Wang Xiaodan, Tang Jie, Waller Jennifer L, Eaton Douglas C, Marrero Mario B: Angiotensin II activation of the JAK/STAT pathway in mesangial cells is altered by high glucose, Kidney Int Suppl 61, 1605-16, 2002 |
| 大鼠 | SD大鼠, 1 周 | Cortical collection duct | Collagenase Type 2: 0.15% | DMEM/Ham's F12 | Valencia L., Bidet M., Martial S., Sanchez E., Melendez E., Tauc M., Poujeol C., Martin D., Namorado M., Reyes J., and Poujeol P.: Nifedipine-activated Ca(2+) Permeability in Newborn Rat Cortical Collecting Duct Cells in Primary Culture, Am J Physiol Cell Physiol280(5), C1193, 2001 |
| 大鼠 | SD大鼠, 250-300克 | Microvessels | Collagenase Type 2: 0.1% | PSS | Li, N., Teggatz, E., Li, P., Allaire, R., and Zou, A.: Formation and Actions of Cyclic ADP-Ribose in Renal Microvessels, Microvasc Res 60, 149, 2000 |
| 大鼠 | SD大鼠, 250-300 克 | Renal | Collagenase Type 2: 0.2% | see reference | Mattson D., and Wu F.: Nitric Oxide Synthase Activity and Isoforms in Rat Renal Vasculature, Hypertension 35, 337, 2000 |
| 大鼠 | SD大鼠, 雄性, 5-6 周 | Renal tubules | Collagenase: 0.1% | see reference | Miyata, N., Park, F., Li, X., and Cowley, A.: Distribution of Angiotensin AT1 and AT2 Receptor Subtypes in the Rat Kidney, Am J Physiol 277, F437, 1999 |
| 大鼠 | SD大鼠, 225-250 g | Glomerular mesangial cells | Collagenase Type 1: 250 u/ml | DMEM | Amiri F, Garcia R: Regulation of angiotensin II receptors and PKC isoforms by glucose in rat mesangial cells, Am J Physiol 276, F691-9, 1999 |
| 大鼠 | SD大鼠, 雄性 | Proximal tubule suspensions | Collagenase Type 4: 0.1% | HEPES buffer | Eitle E, Hiranyachattada S, Wang H, Harris PJ: Inhibition of proximal tubular fluid absorption by nitric oxide and atrial natriuretic peptide in rat kidney, Am J Physiol 274, C1075-80, 1998 |
| Pronase E: 2 u/ml | |||||
| 大鼠 | 大鼠, SD, 雄性 | Nephron segments | Collagenase Type 2: 0.05% | Eagle's MEM | Schafer, J., Watkins, M., Li, L., Herter, P., Haxelmans, S., and Schlatter, E.: A Simplified Method for Isolation of Large Numbers of Defined Nephron Segments, Am J Physiol 273, F650, 1997 |
| Deoxyribonuclease I: 50 u/ml | |||||
| 大鼠 | 大鼠, 12-周大 | Renal | Collagenase: 0.1% | DMEM | Ishikawa, S., Kusaka, I., Higashiyama, M., Nagasaka, S., Saito, T., Honda, K., and Saito, T.: Cellular Signaling and Proliferative Action of AVP in Mesangium of SHR: Effect of Low Density Lipoprotein, Nat Neurosci 50, 1506, 1996 |
| 大鼠 | 大鼠, 雄性, 150-200 克 | Epithelial | Protease: 0.1% | HBSS/CMF | Elliget, K., and Trump, B.: Primary Cultures of Normal Rat Kidney Proximal Tubule Epithelial Cells for Studies of Renal Cell Injury, In Vitro Cell Dev Biol 27, 739, 1991 |
| Proximal tubule | |||||
| 大鼠 | 大鼠, 成年的 | Inner medullary collecting duct | Collagenase Type 2: 0.1% | PBS | Brion, L., Schwartz, J., Lachman, H., Zavilowitz, B., and Schwartz, G.: Development of H+ Secretion by Cultured Renal Inner Medullary Collecting Duct Cells, Am J Physiol 257, F486, 1989 |
| Papillae | |||||
| 大鼠 | Wistar大鼠, 雌性, 100-150 克 | Glomerular mesangial | Collagenase: 0.025% | RPMI 1640 | Wang, J., Kester, M., and Dunn, M.: The Effects of Endotoxin on Plaet-Activating Factor Synthesis in Cultured Rat Glomerular Mesangila Cells, Biochim Biophys Acta 969, 217, 1988 |
| 大鼠 | Wistar大鼠, 雄性, 150 - 200 克 | Renal target | Collagenase: 1.0% (also 0.7%) | Eagle's MEM | Barlet-Bas, C., Khadouri, C., Marsey, S., and Doucet, A.: Sodium-Independent In VitroInduction of Na+,K+-ATPase by Aldosterone in Renal Target Cells: Permissive Effect of Triiodothyronine, Proc Natl Acad Sci U S A 85, 1707, 1988 |
| 大鼠 | Wistar大鼠, 雄性, 成年的 | Proximal tubules | Collagenase: 0.2% | Krebs-Henseleit buffer | Gesek, F., Wolff, D., and Strandhov, J.: Improved Separation Method for Rat Proximal and Distal Renal Tubules, Am J Physiol 253, F358, 1987 |
| 大鼠 | Wistar大鼠, 300-400克 | Proximal tubules | Collagenase: 0.15 % | Krebs Henseleit solution | Vinay, P., Gougoux, A., and Lemieux, G.: Isolation of a Pure Suspension of Rat Proximal Tubules, Am J Physiol 241, F403, 1981 |
| 大鼠 | 大鼠 | Fibroblasts | Trypsin: up to 0.25% | Dulbecco-Vogt MEM | Wallach, D., Anderson, W., and Pastan, I.: Activation of Adenylate Cyclase in Cultured Fibroblasts by Trypsin, J Biol Chem 253, 24, 1978 |
| Kidney |
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