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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。
胰腺組織消化手冊
| 物種 | 詳細來源 | 細胞名稱 | 消化用酶 | 培養基 | 參考文獻 |
| 牛 | 牛 | Duct epithelial | Collagenase: 0.1% | HEPES | Cotton, C., and Al-Nakkash, L.: Isolation and Culture of Bovine Pancreatic Duct Epithelial cells, Am J Physiol 272, G1328, 1997 |
| 牛 | 牛 (歐洲牛) | Ductal | Neutral Protease: 0.05% | EBSS | Sato, T., Sato, M., Hudson, E., and Jones, R.: Characterization of Bovine Pancreatic Ductal Cells Isolated by a Perfusion-Digestion Technique, In Vitro 19, 651, 1983 |
| 牛 | 牛 | Plaets | Trypsin: | (see reference) | Stiles, G., and Lefkowitz, R.: Hormone-Sensitive Adneylate Cyclase, J Biol Chem 257 (11), 6287, 1982 |
| 犬 | 犬 | Buccal ganglia; SLT muscle | Trypsin: 0.2% | DMEM | Zoran, M., Doyle, R. and Haydon, P.: Target Contact Regulates the Calcium Responsiveness of the Secretory Machinery During Synaptogenesis, Neuron 6, 145, 1991 |
| 犬 | 犬, 成年雜種, 15-25 公斤 | Islets | Collagenase Type 4: 600-1100 u/ml | RPMI 1640 | Noel, J., Rabinovitch, A., Olson, L., Kyriakides, G., Miller, J., and Mintz, D.: A Method for Large-Scale, High-Yield Isolation of Canine Pancreatic Islets of Langerhans, Metabolism 31 (2), 184, 1982 |
| Deoxyribonuclease I: 10 ug/ml | |||||
| 魚 | 魚,絲鱸, 3個月大 0.5 克 | Islets | Collagenase: 0.12 - 0.46 u/ml | RPMI 1640 | Schrezenmeir, J., Laue, C., Sternheim,E., Wolbert, K., Darquy, S., Chicheportiche, D., Kirchgessner, J., and Reach, G.: Long-Term Function of Single-Cell Preparations of Piscine Principal Islets in Hollow Fibers, Transplant Proc 24 (6), 2941, 1992 |
| 豚鼠 | 豚鼠 | Acinar | Collagenase Type 3: 60 u/ml | Kreb's Ringer | Schultz, G., Sarras, Jr, M., Gunther, G., Hull, B., Alicea, H., Gorelick, F., and Jamieson, J.: Guinea Pig Pancreatic Acini Prepared with Purified collagenase, Exp Cell Res 130, 49, 1980 |
| 豚鼠 | Hartley豚鼠, 白化變種, 雄性, 350-400 克 | Acinar | Soybean Trypsin Inhibitor: 0.01% | Kreb's Ringer | Gardner, J., Conlon, T., Klaeveman, H., Adams, T., and Ondetti, M.: Action of Cholecystokinin and Cholinergic Agents on Calcium Transport in Isolated Pancreatic Acinar Cells, J Clin Invest 56, 366, 1975 |
| 豚鼠 | 豚鼠 | Exocrine | Hyaluronidase: 0.15% - 0.2% | Kreb's Ringer | Amsterdam, J., and Jamieson, J.: Structural and Functional Characterization of Isolated Pancreatic Exocrine Cells, Proc Natl Acad Sci U S A 69 (10), 3028, 1972 |
| 馬 | 馬 | Islets | Collagenase Type 4: 1.3% - 2.0% | HBSS | Feldman, J., and Chapman, B.: Preparation of Islets of Langerhans from Rabbits and Hamsters by the Collagenase Digestion Technique, Acta Diabetol 12, 208, 1975 |
| 人類 | 人類 | Acinar | CLSPA: 200 u/ml | See Reference | Cane, M., Sutton, R. and Criddle, D.: Isolation of Human Pancreatic Acinar Cells From Rat and Human Pancreas, Pancreapedia 20, 1, 2011 |
| Soybean Trypsin Inhibitor: 0.01% | |||||
| 人類 | 人類 | Pancreatic cancer stem cells | Collagenase Type 4: 200 u/ml | medium 199 | Li, C., Heidt, D., Dalerba, P., Burant, C., Zhang, L., Adsay, V., Wicha, M., Clarke, M. and Simeone, D.: Identification of Pancreatic Cancer Stem Cells., Cancer Res 67, 1030, 2007 |
| 人類 | 人類, 25-55 歲大 | Islets | Collagenase: 0.4% | HBSS | Contractor, H., Johnson, P., Chadwick, D., Robertson, G., and London, N.: The Effect of UW Solution and Its Components on the Collagenase Digestion of Human and Porcine Pancreas, Cell Transplant 4 (6), 615, 1995 |
| (包括 豬) | |||||
| 人類 | 人類 | Islets | Collagenase: 0.2% | Eurocollins solution | Watt, P., Mullen, Y., Benhamou, P., Hober, C., Nomura, Y., Watanabe, Y., Passaro, E., Zinner, M., and Brunicardi, F.: Simplified Semiautomated Method For Isolating Islets From the Human Pancreas, Transplant Proc 26 (2), 582, 1994 |
| Deoxyribonuclease I: 200 u/ml | |||||
| 人類 | 人類, 成人器官捐獻者 | Islets | Collagenase: 0.6% | Eurocollins solution | Warnock, G., Rajotte, R., Evans, M., Ellis, D., DeGroot, T., and Dawidson, I.: Isolation of Islets of Langerhans Following Cold Storage of Human Pancreas, Transplant Proc XIX (4), 3466, 1987 |
| 人類 | 人類 | Islets | Collagenase Type 4: 0.8% | HBSS | Izumi, R., Konishi, K., Ueno, K., Shimizu, K., Hirosawa, H., Takahashi, N., and Miyazaki, I.: Isolation of Human Pancreatic Islets from Cryopreserved Pancreas,Transplant Proc XVII, 383, 1985 |
| 人類 | 人類 | Islets | Collagenase (1 or 4): 0.60% | HBSS | Gray, D., McShane, P., Grant, A., and Morris, P.: A Method for Isolation of Islets of Langerhans from the Human Pancreas, Diabetes 33, 1055, 1984 |
| 人類 | 人類, 嬰兒, a克e 1 天-1 歲 | Islets | Collagenase: 170-210 u/ml | HBSS | Sutherland, D., Matas, A., Steffes, M., and Najarian, J.: Infant Human Pancreas: A Potential Source of Islet Tissue for Transplantation, Diabetes 25 (12), 1123, 1976 |
| 猴 | 猴, 3-5 公斤 | Islets | Hyaluronidase: 0.05% | HBSS | Scharp, D., Murphy, J., Newton, W., Ballinger, W., and Lacy, P.: Application of an Improved Isolation Technique for Islet Transplantation in Primates and Rats,Transplant Proc 7, 739, 1975 |
| 小鼠 | 小鼠 | Islets | Collagenase: 0.1-0.25% | HBSS | Koh, D., Moody, M. and Jo, J.: Collection of Islets of Langerhans using an Equilibrium Method., Biotechniques 55, 34-7, 2013 |
| 小鼠 | 小鼠, 8-12 周 | Pancreas organoid | Collagenase: 0.012% | DMEM | Huch, M., Bonfanti, P., Boj, S., Sato, T., Loomans, C., Van de Wetering, M., Sojoodi, M., Li, V., Schuijers, J., Gracanin, A., Ringnalda, F., Begthel, H., Hamer, K., Mulder, J., Van Es, J. and De Koning, E.: Unlimited In Vitro Expansion of Adult Bi-Potent Pancreas Progenitors Through the Lgr5/R-Spondin Axis., EMBO J 32, 2708-21, 2013 |
| Neutral Protease: 0.012% | |||||
| 小鼠 | 小鼠, 胚胎 | Pancreatic progenitor | Neutral Protease: 0.125% | DMEM | Greggio, C., De Franceschi, F., Figueiredo-Larsen, M., Gobaa, S., Ranga, A., Semb, H., Lutolf, M. and Grapin-Botton, A.: Artificial Three-Dimensional Niches Deconstruct Pancreas Development In Vitro., Development 140, 4452-62, 2013 |
| 小鼠 | 小鼠 | Islets | Collagenase: 0.2% | RPMI 1640 | Bertera, S., Balamurugan, A., Bottino, R., He, J. and Trucco, M.: Increased Yield and Improved Transplantation Outcome of Mouse Islets with Bovine Serum Albumin., J Transplant 2012, 856386, 2012 |
| 小鼠 | 小鼠, 10 周 | Islets | Collagenase: 0.2% | RPMI 1640 | Kobayashi, T., Yamaguchi, T., Hamanaka, S., Kato-Itoh, M., Yamazaki, Y., Ibata, M., Sato, H., Lee, Y., Usui, J., Knisely, A., Hirabayashi, M. and Nakauchi, H.: Generation of Rat Pancreas in Mouse by Interspecific Blastocyst Injection of Pluripotent Stem Cells., Cell142, 787, 2010 |
| 小鼠 | 小鼠, 6-8 個月 | Islets | Collagenase Type 4: 0.1% | HBSS | Taguchi, Y., Tasaki, Y., Terakado, K., Kobayashi, K., Machida, T. and Kobayashi, T.:Impaired Insulin Secretion from the Pancreatic Islets of Hypothyroidal Growth-Retarded Mice., J Endocrinol 206, 195-204, 2010 |
| 小鼠 | 小鼠, 成年的, 雄性 | Ancinar | CLSPA: see reference | DMEM | Ji, B., Gaiser, S., Chen, X., Ernst, S. and Logsdon, C.: Intracellular Trypsin Induces Pancreatic Acinar Cell Death but not NF-KappaB Activation., J Biol Chem 284, 17488, 2009 |
| Soybean Trypsin Inhibitor: 0.001% | |||||
| 小鼠 | 小鼠, 3-4 周, 20-24 克 | Islets | Collagenase Type 4: 0.2% | RPMI 1540 | Huang, H., Xie, Q., Kang, M., Zhang, B., Zhang, H., Chen, J., Zhai, C., Yang, D., Jiang, B. and Wu, Y.: Labeling Transplanted Mice Islet with Polyvinylpyrrolidone Coated Superparamagnetic Iron Oxide Nanoparticles for In Vivo Detection By Magnetic Resonance Imaging., Nanotechnology 20, 365101, 2009 |
| 小鼠 | 小鼠 | Islets | Collagenase: 1,000 u/ml | RPMI 1640 | Li, D., Yuan, Y., Tu, H., Liang, Q. and Dai, L.: A Protocol for Islet Isolation from Mouse Pancreas., Nat Protoc 4, 1649, 2009 |
| 小鼠 | 小鼠 | Islets | Collagenase: 0.14% | RPMI 1640 | Carter, J., Dula, S., Corbin, K., Wu, R. and Nunemaker, C.: A Practical Guide to Rodent Islet Isolation and Assessment., Biol Proced Online 11, 3, 2009 |
| 小鼠 | 小鼠 | Islets | Collagenase: 0.03-0.08% | RPMI 1640 | Szot, G., Koudria, P. and Bluestone, J.: Murine Pancreatic Islet Isolation., J Vis Exp 7, 255, 2007 |
| 小鼠 | 小鼠, 6-10 個月 | Pancreatic ductal | CLSPA: 50 u/ml | DMEM | Wang, Y., Soyombo, A., Shcheynikov, N., Zeng, W., Dorwart, M., Marino, C., Thomas, P. and Muallem, S.: Slc26a6 Regulates CFTR Activity In Vivo to Determine Pancreatic Duct HCO3- Secretion: Relevance to Cystic Fibrosis., EMBO J 25, 5049, 2006 |
| Hyaluronidase: 400 u/ml | |||||
| Soybean Trypsin Inhibitor: 0.02% | |||||
| 小鼠 | 小鼠, 成年的 | Acinar | CLSPA: 200 u/ml | see reference | Voronina, S., Barrow, S., Gerasimenko, O., Petersen, O. and Tepikin, A.: Effects of Secretagogues and Bile Acids on Mitochondrial Membrane Potential of Pancreatic Acinar Cells: Comparison of Different Modes of Evaluating Delta Psi., J Biol Chem 279, 27327, 2004 |
| 小鼠 | 小鼠, 11-12 周 | Islets | Collagenase Type 4: 0.2% | RPMI | Astrof, S., Crowley, D., George, E., Fukuda, T., Sekiguchi, K., Hanahan, D. and Hynes, R.: Direct Test of Potential Roles of EIIIA and EIIIB Alternatively Spliced Segments of Fibronectin in Physiological and Tumor Angiogenesis., Mol Cell Biol 24, 8662-70, 2004 |
| 小鼠 | 小鼠 | Pancreatic islet | Collagenase Type 4: 0.2% | HBSS | Haefliger Jacques-Antoine, Tawadros Thomas, Meylan Laure, Gurun SabineLe, Roehrich Marc-Estienne, Martin David, Thorens Bernard, Waeber Gerard: The scaffold protein IB1/JIP-1 is a critical mediator of cytokine-induced apoptosis in pancreatic beta cells, J Cell Sci 116, 1463-9, 2003 |
| 小鼠 | 小鼠 | Islets | Collagenase Type 4: 0.2% | HBSS | Wu Yulian, Han Bing, Luo Hongyu, Roduit Raphael, Salcedo TheodoraW, Moore PaulA, Zhang Jun, Wu Jiangping: DcR3/TR6 effectively prevents islet primary nonfunction after transplantation, Diabetes 52, 2279-86, 2003 |
| 小鼠 | 小鼠, BALB/c, 6-8 周-old, either sex | Acinar | Collagenase: 0.1% | Waymouth's MB | Kurup, S., and Bhonde, R.: Analysis and Optimization of Nutritional Set-up for Murine Pancreatic Acinar Cells, JOP 3 (1), 8, 2002 |
| 小鼠 | 小鼠 | Islets | Collagenase Type 2: 0.2% | CF Medium | Koster, J., Marshall, B., Ensor, N., Corbett, J., and Nichols, C.: Targeted Overativity of Cell KATP Channels Induces Profound Neonatal Diabetes, Cell 100, 645, 2000 |
| 小鼠 | 小鼠 | Pancreatic islets | Collagenase Type 4: 0.4% | Gey's BSS | Strowski M, Parmar R, Blake A, Schaeffer J: Somatostatin inhibits insulin and glucagon secretion via two receptors subtypes: an in vitro study of pancreatic islets from somatostatin receptor 2 knockout mice, Endocrinology 141, 111-7, 2000 |
| 小鼠 | 小鼠 | Acinar cells and acini | Collagenase Type 1: see reference | see reference | Toivola, D., Ku, N., Ghori, N., Lowe, A., Michie, S. and Omary, M.: Effects of Keratin Filament Disruption on Exocrine Pancreas-Stimulated Secretion and Susceptibility to Injury., Exp Cell Res 255, 156, 2000 |
| CLSPA: see reference | |||||
| 小鼠 | 小鼠, 白話病變, 25 克 | Acinar | CLSPA: see reference | see reference | Fogarty, K., Kidd, J., Tuft, R. and Thorn, P.: A Bimodal Pattern of InsP(3)-Evoked Elementary Ca(2+) Signals in Pancreatic Acinar Cells., Biophys J 78, 2298, 2000 |
| 小鼠 | 小鼠 | Duct | Papain: 25 u/ml | DMEM /F-12 | Githens, S: Pancreatic Duct Epithelial Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:12.1, 1995 |
| 小鼠 | 小鼠 | Acinar | Collagenase: 100 u/ml | HEPES | Jauch, P., Peterson, O., and Lauger, P.: Electrogenic Properties of the Na-Alanine Cotransporter in Acinar Cells, J Membr Biol 94, 99, 1986 |
| 小鼠 | 小鼠 (C57BL/6J-ob/ob), 9-12 周, 雄性 | Islets | Hyaluronidase: 0.5% | Kreb's Ringer bicarbonate buffer | Dalpe-Scott, M., Heick, H., and Begin-Heick, N.: Secretion in the Obese (ob/ob) Mouse. The Effect of Oxytetracycline on Insulin Release, Diabetes 32, 932, 1983 |
| 小鼠 | 小鼠, 雄性, 18-24 克 | Acinar | CLSPA: 70-90 u/ml | Krebs-Henseleit | Burnham DB, Williams JA: Effects of carbachol, cholecystokinin, and insulin on protein phosphorylation in isolated pancreatic acini, J Biol Chem 257, 10523-8, 1982 |
| Soybean Trypsin Inhibitor: 0.01% | |||||
| 小鼠 | 小鼠, 7-10 周 | Islets | Collagenase: see reference | DMEM | Yesil, P, Michel, M., Chwalek, K., Pedack, S., Jany, C., Ludwig, B., Bornstein, S. and Lammert, E.: A New Collagenase Blend Increases the Number of Islets Isolated from Mouse Pancreas., Islets 1, 185, |
| 豬 | 豬, 少年的 | Islets | Collagenase: see reference | Univ of Wisconsin solution | Van der Burg Michael P M, Graham John M: Iodixanol Density Gradient Preparation in University of Wisconsin solution for porcine islet purification, ScientificWorldJournal 3, 1154-9, 2003 |
| 豬 | 豬, 3 個月, 15-20 公斤 | Acinar | Collagenase Type 3: 200 u/ml | RPMI 1640 | Zhao, X., Han, J., and Tang, C.: Primary Culture of Porcine Pancreatic Acinar Cells, JOP2 (2), 78, 2001 |
| 豬 | 豬, 1-3 歲, 1.5-2.0 公斤, 任一性別 | Islets | Collagenase: 0.25% | HBSS | Korbutt, G., Elliott, J., Ao, Z., Smith, D., Warnock, G., and Rajotte, R.: Large Scale Isolation, Growth, and Function of Porcine Neonatal Islet Cells, J Clin Invest 97 (9), 2119, 1996 |
| 豬 | 豬, 雌性 | Islets | Collagenase: 0.1% | HBSS | Brandhorst, D., Brandhorst, H., Hering, B., Federlin, K., and Bretzel, R.: Islet Isolation from the Pancreas of Large Mammals and Humans: 10 歲s of Experience, Exp Clin Endocrinol 103, 3, 1995 |
| 豬 | 豬 | Islets | Collagenase: 0.1%-0.2% | HBSS | Johnson, P., van Suylichem, P., Roberts, D., Vos-Scheperkeuter, G., White, S., van Schlifgaarde, R., London, N.: Design of a Simple, in vitro Method for Evaluation of the Efficiency of Crude Clostridium histolyticum Collagenase and its Components for Porcine Islet Isolation, Xenotransplantation 2, 165, 1995 |
| 豬 | 豬, <2 歲, 200-250 公斤 | Islets | Collagenase: 0.1% | HBSS | Heiser, A., Ulrichs, K., and Muller-Ruchholtz, W.: Isolation of Porcine Pancreatic Islets: Low Trypsin Activity During the Isolation Procedure Guarantees Reproducible High Islet Yields, J Clin Lab Anal 8, 407, 1994 |
| 豬 | 豬, 10-36 個月s, 200-300 公斤 | Islets | Collagenase: 0.2% | HBSS | Ricordi, C., Socci, C., Davalli, A., Staudacher, C., Baro, P., Vertova, A., Sassi, I., Gavazzi, F., Pozza, G., and Di Carlo, V.: Isolation of the Elusive Pig Islet, Surgery 107 (6), 688, 1990 |
| 豬 | 豬 | Acinar | Collagenase: 100 u/ml | Saline | Iwatsuki, N., and Peterson, O.: Action of Tetraethylammonium on Calcium-Activated Potassim Channels in Pig Pancreatic Acinar Cells Studied by Patch-Clamp Single-Channel and Whole-Cell Current Recording, J Membr Biol 86, 139, 1985 |
| 兔 | 兔, 新西蘭白, 雄性/Fe雄性, 2-3 公斤 | Acinar | Hyaluronidase: 0.2% | Kreb's Ringer bicarbonate buffer | Renckens, B., Schrijen, J., Swarts, H., DePont, J., and Bonting, S.: Role of Calcium in Exocrine Pancreatic Secretion. IV. Calcium Movements in Isolated Acinar Cells of Rabbit Pancreas, Biochim Biophys Acta 544, 338, 1978 |
| 兔 | 兔, 包括馬 | Islets | Collagenase Type 4: 1.3% - 2.0% | HBSS | Feldman, J., and Chapman, B.: Preparation of Islets of Langerhans from Rabbits and Hamsters by the Collagenase Digestion Technique, Acta Diabetol 12, 208, 1975 |
| 大鼠 | Wistar大鼠, 300克 | Islets | Collagenase Type 4: 0.2% | CMRL 1066 | Verga Falzacappa, C., Mangialardo, C., Raffa, S., Mancuso, A., Piergrossi, P., Moriggi, G., Piro, S., Stigliano, A., Torrisi, M., Brunetti, E., Toscano, V. and Misiti, S.: The Thyroid Hormone T3 Improves Function and Survival of Rat Pancreatic Islets During In Vitro Culture., Islets 2, 96, 2010 |
| 大鼠 | 大鼠 | Acinar | CLSPA: 100 u/ml | DMEM | Williams, J.: Isolation of Rodent Pancreatic Acinar Cells and Acini by Collagenase Digestion, Pancreapedia , , 2010 |
| 大鼠 | 大鼠, 成年的 | Islets | Collagenase Type 1: 450 u/ml | Leibowitz L-15 | Williams, J., Huang, H., Kover, K., Moore, W., Berkland, C., Singh, M., Smirnova, I., MacGregor, R. and Stehno-Bit, L.: Reduction of Diffusion Barriers in Isolated Rat Islets Improves Survival, but not Insulin Secretion or Transplantation Outcome., Organogenesis6, 115, 2010 |
| 大鼠 | 大鼠, 雄性, 7-11 周 | Islets | Collagenase Type 4: see reference | RPMI 1640 | Getty-Kaushik, L., Richard, A., Deeney, J., Shirihai, O. and Corkey, B.: The CB1 Antagonist Rimonabant Decreases Insulin Hypersecretion in Rat Pancreatic Islets, Obesity17, 1856, 2009 |
| 大鼠 | 大鼠 | Islets | Collagenase Type 1: 450 u/ml | RPMI 1640 | MacGregor, R., Williams, S., Tong, P., Kover, K., Moore, W. and Stehno-Bit, L.:Small Rat Islets are Superior to Large Islets in In Vitro Function and in Transplantation Outcomes., Am J Physiol/Endo 290, E771, 2006 |
| 大鼠 | Wistar大鼠, 雄性, 250-400克 | Pancreatic islets | Collagenase: 0.75% | RPMI 1640 | Tian XH, Xue WJ, Ding XM, Pang XL, Teng Y, Tian PX, and Feng XS: Small intestinal submucosa improves islet survival and function during in vitro culture, World J Gastroenterol 11, 7378, 2005 |
| 大鼠 | 大鼠, 250-350 克 | Pancreatic islet | Collagenase Type 4: 0.2% | HBSS | Haefliger Jacques-Antoine, Tawadros Thomas, Meylan Laure, Gurun SabineLe, Roehrich Marc-Estienne, Martin David, Thorens Bernard, Waeber Gerard: The scaffold protein IB1/JIP-1 is a critical mediator of cytokine-induced apoptosis in pancreatic beta cells, J Cell Sci 116, 1463-9, 2003 |
| 大鼠 | 大鼠 | Pancreatic acini | CLSPA: 30 u/ml | M199 | Blinman TA, Gukovsky I, Mouria M, Zaninovic V, Livingston E, Pandol SJ, Gukovskaya AS: Activation of pancreatic acinar cells on isolation from tissue: cytokine upregulation via p38 MAP kinase, Am J Physiol Cell Physiol 279, C1993-2003, 2000 |
| Collagenase Type 4: 30 u/ml | |||||
| Soybean Trypsin Inhibitor: 0.01% | |||||
| 大鼠 | SD大鼠, 雄性 | Ancinar | CLSPA: see reference | DMEM | Ji B, Kopin AS, Logsdon CD: Species differences between rat and mouse CCKA receptors determine the divergent acinar cell response to the cholecystokinin analog JMV-180, J Biol Chem 275, 19115-20, 2000 |
| Soybean Trypsin Inhibitor: 0.01% | |||||
| 大鼠 | 大鼠 | Islets | Collagenase: 126 - 196 u/ml | HBSS | Verspohl, E., and Wienecke, A.: The Role of Protein Kinase C in the Desensitization of Rat Pancreatic Islets to Chrolinergic stimulation, J Endocrinol 159, 287, 1998 |
| 大鼠 | 大鼠 | Islets | Collagenase: 0.5 - 0.9% | HBSS | Takaki, R and Ono J: Culture of Pancreatic Islet Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:11.1, 1995 |
| 大鼠 | 大鼠 | Duct | Papain: 25 u/ml | DMEM /F-12 | Githens, S: Pancreatic Duct Epithelial Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:12.1, 1995 |
| 大鼠 | SD大鼠, 雄性, 40 - 100 克 | Acinar | Hyaluronidase: 462 u/ml | Ham's F12 | Hirschi, K., Kenny, S., Justice, J., Brannon, P.: Effects of Secretin And Caerulein On Enzymes Of Cultured Pancreatic Acinar Cells, In Vitro Cell Dev Biol 27, 660, 1991 |
| 大鼠 | SD大鼠, 雄性, 100 - 150 克 | Parotid acinar | Trypsin: 0.001% | F12 medium | Yeh, C., Mertz, P., Oliver, C., Baum, B., and Kousvelari, E.: Cellular Characteristics of Long-Term Cultured Rat Parotid Acinar Cells, In Vitro Cell Dev Biol 27, 707, 1991 |
| 大鼠 | Wistar大鼠, 雄性 | Parotid acinar | Trypsin: 0.02% | Solution B | Foskett, J., Roifman, C. and Wong, D.: Activation of Calcium Oscillations by Thapsigargin in Parotid Acinar Cells, J Biol Chem 266, 2778, 1991 |
| (See reference) | |||||
| 大鼠 | Wistar大鼠, 雄性, 230 - 270 克 | Islets | Collagenase: 0.1% - 0.2% | HBSS | Ohzato, H., Gotoh, M., Monden, M., Dono, K., Kanai, T., and Mori, T.: Improvement in the Islet Yield From a Cold-Preserved Pancreas by Pancreatic Ductal Collagenase Distention at the Time of Harvesting, Transplantation 51, 566, 1991 |
| (包括小鼠, 6 - 8 周大) | |||||
| 大鼠 | SD大鼠, 任一性別, 6-20 周大 | Interlobular ducts | Papain: 25 u/ml | DMEM/Ham's F-12 | Githens, S., Schexnayder, J., Desai, K., and Patke, C.: Rat Pancreatic Interlobular Duct Epithelium: Isolation and Culture in Collagen Gel, In Vitro Cell Dev Biol 25 (8), 679, 1989 |
| 大鼠 | SD大鼠, 雄性, 150 - 200 克 | Acinar | Soybean Trypsin Inhibitor: 0.01% | HEPES | Menozzi, D., Sato, S., Jensen, R., and Gardner, J.: Cyclic GMP Does Not Inhibit Protein Kinase C-Mediated Enzyme Secretion in Rat Pancreatic Acini, J Biol Chem 264, 995, 1989 |
| 大鼠 | Wistar大鼠, 雄性, 250 - 350 克 | Acinar, parotid | Hyaluronidase: 0.015% | Earle's MEM | Melvin, J., Kawaguchi, M., Baum, B., and Turner, R.: A Muscarinic Agonist-Stimulated Chloride Efflux Pathway is Associated With Fluid Secretion in Rat Parotid Acinar Cells,Biochem Biophys Res Commun 145, 754, 1987 |
| 大鼠 | SD大鼠, 雄性, 50 - 125 克 | Acinar | Trypsin: 0.01% | HBSS, CMF | Oliver, C., Waters, J., Tolbert, C., and Kleinman, H.: Growth of Exocrine Acinar Cells on a Reconstituted Basement Membrane Gel, In Vitro Cell Dev Biol 23, 465, 1987 |
| Exorbital lacrimal, parotid, pancreas | |||||
| 大鼠 | Fischer-344大鼠, 任一性別, 120-150 克 | Epithelial | Trypsin: 0.1% | Ham's F-12/HBSS | Tsao, M., and Duguid, W.: Establishment of Propagable Epithelial Cell Lines From Normal Adult Rat Pancreas, Exp Cell Res 168, 365, 1987 |
| (see reference) | |||||
| 大鼠 | 大鼠, 200-350 克 | Islets | Collagenase Type 4: 0.2% | HBSS | Gotoh, M., Maki, T., Satomi, S., Porter, J., Bonner-Weir, S., O'Hara, C., and Monaco, A.: Reproducible High Yield of Rat Islets By Stationary In Vitro Digestion Following Pancreatic Ductal or Portal Venous Collagenase Injection, Transplantation 43 (5), 725, 1987 |
| 大鼠 | SD大鼠, 雄性, 42 - 48 天, 175 - 200 克 | Acinar, submandibular gland | Hyaluronidase: 0.1 % | HBSS, CF | Quissell, D., Redman, R., and Mark, M.: Short-Term Primary Culture of Acinar-Intercalated Duct Complexes From Rat Submandibular Glands, In Vitro Cell Dev Biol 22, 469, 1986 |
| 大鼠 | 暗條紋和劉易斯大鼠, 供血者, 成年的 | Islets | Collagenase Type 1: 0.3% | Hank's solution | Sutton, R., Peters, M., McShane, P., Gray, D., and Morris, P.: An Improved Method for the Isolation of Islets of Langerhans From the Adult Rat Pancreas, Transplant Proc XVII (6), 1819, 1986 |
| 大鼠 | SD大鼠, 雄性, 50-75 克 | Acinar | Hyaluronidase: 0.1% | HBSS | Brannon, P., Orrison, B., and Kretchmer, N.: Primary Cultures of Rat Pancreatic Acinar Cells in Serum-Free Medium, In Vitro Cell Dev Biol 21 (1), 6, 1985 |
| (see reference) | |||||
| 大鼠 | SD大鼠, 125 - 350 克 | Duct | Trypsin: 0.01% | HBSS | Githens, S., Holmquist, D., Whelan, J., and Ruby, J.: Ducts of the Rat Pancreas in Agarose Matrix Culture, In Vitro 16, 797, 1980 |
| 大鼠 | 大鼠 | Exocrine | Hyaluronidase: 0.9% | Kreb's Ringer | Schulz, I., Heil, K., Kribben, A., Sachs, G., and Haase, W.: Isolation and Functional Characterization of Cells From the Exocrine Pancreas,Biology of Normal and Cancerous Exocrine Pancreatic Cells,Ribet, A., Pradayrol, L., and Susini, C., Elsevier, , 1980 |
| 大鼠 | Wistar大鼠, 雄性 | Islets | Collagenase Type 4: 1% | Medium 199 | Katada, T., and Ui, M.: Enhanced Insulin Secretion and Cyclic Amp Accumulation in Pancreatic Islets Due to Activation of Native Calcium Ionophores, J Biol Chem 254 (2), 469, 1979 |
| 大鼠 | SD大鼠, 雄性, 250 - 350 克 | Acinar | Hyaluronidase: 0.18% | Kreb's Henseleit bicarbonate buffer | Williams, J., Korc, M., and Dormer, R.: Action of Secretagogues on a New Preparation of Functionally Intact, Isolated Pancreatic Acini, Am J Physiol 235, 517, 1978 |
| (包括小鼠, 小白鼠, 瑞士鼠, 雄性, 20 - 24 克) | |||||
| 大鼠 | Wistar大鼠, 雄性, 300-350 克 | Islets | Collagenase: 0.2% | Kreb's Ringer bicarbonate buffer | Wolters, G., Konijenendijk, W., and Bouman, P.: Effects of Fasting on Insulin Secretion, Islet Glucose Metabolism, and the Cyclic Adenosine 3'5'-Monophosphate Content of Rat Pancreatic Islets In Vitro, Diabetes 26 (6), 530, 1977 |
| 大鼠 | Wistar大鼠, 雄性, 200 - 300 克 | Islets | Collagenase Type 4: 0.5% | HBSS | Shibata, A., Ludvigsen, C., Naber, S., McKaneil M., and Lacy, P.: Standardization of a Digestion-Filtration Method for Isolation of Islets, Diabetes 25, 667, 1976 |
| 大鼠 | 大鼠 | Exocrine | Hyaluronidase: 0.15% | Krebs | Kondo, S., and Schulz, I.: Calcium Ion Uptake in Isolated Pancreas Cells Induced by Secretagogues, Biochim Biophys Acta 419, 76, 1976 |
| 大鼠 | Wistar大鼠-Lewis大鼠, 胎兒 18-20 妊娠日 | Islets | Collagenase Type 4: 0.63% | EBSS | Braaten, J., Jarlfors, U., Smith, D., and Mintz, D.: Purification of Monolayer Cell Cultures of the Endocrine Pancreas, Tissue Cell 7 (4), 747, 1975 |
| (包括新生 3-12 天大) | (see reference) | ||||
| 大鼠 | SD大鼠, 白話病變, 雄性, 150-200 克 | Acinar, parotid | Trypsin: 0.01% | HBSS CMF | Mangos, J., McSherry, N., Butcher, F., Irwin, K., and Barber, T.: Dispersed Rat Parotid Acinar Cells (Morphological, Functional Character), Am J Physiol 229 (3), 560, 1975 |
| Collagenase: 40-50 u/ml | |||||
| Hyaluronidase: 0.10% | |||||
| 大鼠 | 大鼠, 新生的 | Islets | Trypsin: 0.05% | Puck's saline buffered w/ EDTA 0.02% | Leonard, R., Lazarow, A., and Hegre, O.: Pancreatic Islet Transplantation in the Rat,Diabetes 22, 413, 1973 |
| 大鼠 | 大鼠, 雄性, 白話病變, 200-300 克 | Islets | Collagenase: 0.5% | Hanks solution | Lacy, P., Walker, M., and Fink, J.: Perifusion of Isolated Rat Islets in Vitro, Diabetes 21 (10), 987, 1972 |
| 大鼠 | Lewis大鼠, 近親繁殖的 | Islets | Collagenase: 0.5% | HBSS | Ballinger, W., and Lacy, P.: Transplantation of Intact Pancreatic Islets in Rats, Surgery 72 (2), 175, 1972 |
| 大鼠 | Wistar大鼠, 白話病變, 雄性, 400 - 500 克 | Islets | Collagenase Type 4: 1.0% - 1.2% | HBSS | Lacy, P., and Kostianovsky, M.: Method for the Isolation of Intact Islets of Langerhans from the Rat Pancreas, Diabetes 16, 35, 1967 |
| 大鼠 | 大鼠 | Ascites hepatoma | Trypsin: 0.1% | Phosphate buffer (see reference) | Essner, E.: Experiments on an Ascites Hepatoma. I. Enzymatic Digestion and Alkaline Degradation of the Cementing Substance and Separation of Cells, in Tumor Islands, Exp Cell Res 7, 430, 1954 |
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