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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，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)具體實驗?zāi)康男拚桨敢垣@得更好的實驗結(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、孵育時間
通常前四個原因沒辦法選擇，而剩下的原因一般由經(jīng)驗決定以獲取zui適實驗結(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驗數(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)胞收集知識和方法，描述了標(biāo)準(zhǔn)的實驗步驟， 以便于制定一個*的細(xì)胞分離方案，同時列舉了一些相關(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。
胰腺組織消化手冊
| 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 | 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(xiàn)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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