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        1. 上海起發(fā)實驗試劑有限公司

          胰腺組織消化手冊

          時間:2016-2-16閱讀:961

          世界*實驗材料供應商 Worthington Biochemical 正式授權上海起發(fā)為其中國代理, Worthington Biochemical 在一直是行業(yè)的*,一直為廣大科研客戶提供zui為的產品和服務,上海起發(fā)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(yǎng)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(jù)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(yǎng)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(shù)據(jù)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(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, 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

          心臟

          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(yǎng)基參考文獻
          Duct epithelialCollagenase: 0.1%HEPESCotton, C., and Al-Nakkash, L.: Isolation and Culture of Bovine Pancreatic Duct Epithelial cells, Am J Physiol 272, G1328, 1997
          牛 (歐洲牛)DuctalNeutral Protease: 0.05%EBSSSato, 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
          PlaetsTrypsin: (see reference)Stiles, G., and Lefkowitz, R.: Hormone-Sensitive Adneylate Cyclase, J Biol Chem 257 (11), 6287, 1982
          Buccal ganglia; SLT muscleTrypsin: 0.2%DMEMZoran, M., Doyle, R. and Haydon, P.: Target Contact Regulates the Calcium Responsiveness of the Secretory Machinery During Synaptogenesis, Neuron 6, 145, 1991
          犬, 成年雜種, 15-25 公斤IsletsCollagenase Type 4: 600-1100 u/mlRPMI 1640Noel, 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,絲鱸, 3個月大 0.5 克IsletsCollagenase: 0.12 - 0.46 u/mlRPMI 1640Schrezenmeir, 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AcinarCollagenase Type 3: 60 u/mlKreb's RingerSchultz, 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AcinarSoybean Trypsin Inhibitor: 0.01%Kreb's RingerGardner, 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ExocrineHyaluronidase: 0.15% - 0.2%Kreb's RingerAmsterdam, J., and Jamieson, J.: Structural and Functional Characterization of Isolated Pancreatic Exocrine Cells, Proc Natl Acad Sci U S A 69 (10), 3028, 1972
          IsletsCollagenase Type 4: 1.3% - 2.0%HBSSFeldman, 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AcinarCLSPA: 200 u/mlSee ReferenceCane, 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 cellsCollagenase Type 4: 200 u/mlmedium 199Li, 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IsletsCollagenase: 0.4%HBSSContractor, 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IsletsCollagenase: 0.2%Eurocollins solutionWatt, 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IsletsCollagenase: 0.6%Eurocollins solutionWarnock, 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IsletsCollagenase Type 4: 0.8%HBSSIzumi, 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IsletsCollagenase (1 or 4): 0.60%HBSSGray, 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IsletsCollagenase: 170-210 u/mlHBSSSutherland, 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, 3-5 公斤IsletsHyaluronidase: 0.05%HBSSScharp, 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IsletsCollagenase: 0.1-0.25%HBSSKoh, 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 organoidCollagenase: 0.012%DMEMHuch, 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 progenitorNeutral Protease: 0.125%DMEMGreggio, 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IsletsCollagenase: 0.2%RPMI 1640Bertera, 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IsletsCollagenase: 0.2%RPMI 1640Kobayashi, 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IsletsCollagenase Type 4: 0.1%HBSSTaguchi, 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AncinarCLSPA: see referenceDMEMJi, 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IsletsCollagenase Type 4: 0.2%RPMI 1540Huang, 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IsletsCollagenase: 1,000 u/mlRPMI 1640Li, 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IsletsCollagenase: 0.14%RPMI 1640Carter, 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IsletsCollagenase: 0.03-0.08%RPMI 1640Szot, 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 ductalCLSPA: 50 u/mlDMEMWang, 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AcinarCLSPA: 200 u/mlsee referenceVoronina, 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IsletsCollagenase Type 4: 0.2%RPMIAstrof, 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 isletCollagenase Type 4: 0.2%HBSSHaefliger 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IsletsCollagenase Type 4: 0.2%HBSSWu 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 sexAcinarCollagenase: 0.1%Waymouth's MBKurup, 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IsletsCollagenase Type 2: 0.2%CF MediumKoster, 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 isletsCollagenase Type 4: 0.4%Gey's BSSStrowski 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 aciniCollagenase Type 1: see referencesee referenceToivola, 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AcinarCLSPA: see referencesee referenceFogarty, 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DuctPapain: 25 u/mlDMEM /F-12Githens, 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AcinarCollagenase: 100 u/mlHEPESJauch, 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IsletsHyaluronidase: 0.5%Kreb's Ringer bicarbonate bufferDalpe-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AcinarCLSPA: 70-90 u/mlKrebs-HenseleitBurnham 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IsletsCollagenase: see referenceDMEMYesil, 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, 少年的IsletsCollagenase: see referenceUniv of Wisconsin solutionVan 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, 3 個月, 15-20 公斤AcinarCollagenase Type 3: 200 u/mlRPMI 1640Zhao, X., Han, J., and Tang, C.: Primary Culture of Porcine Pancreatic Acinar Cells, JOP2 (2), 78, 2001
          豬, 1-3 歲, 1.5-2.0 公斤, 任一性別IsletsCollagenase: 0.25%HBSSKorbutt, 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, 雌性IsletsCollagenase: 0.1%HBSSBrandhorst, 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
          IsletsCollagenase: 0.1%-0.2%HBSSJohnson, 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, <2 歲, 200-250 公斤IsletsCollagenase: 0.1%HBSSHeiser, 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, 10-36 個月s, 200-300 公斤IsletsCollagenase: 0.2%HBSSRicordi, 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
          AcinarCollagenase: 100 u/mlSalineIwatsuki, 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, 新西蘭白, 雄性/Fe雄性, 2-3 公斤AcinarHyaluronidase: 0.2%Kreb's Ringer bicarbonate bufferRenckens, 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, 包括馬IsletsCollagenase Type 4: 1.3% - 2.0%HBSSFeldman, 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IsletsCollagenase Type 4: 0.2%CMRL 1066Verga 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AcinarCLSPA: 100 u/mlDMEMWilliams, J.: Isolation of Rodent Pancreatic Acinar Cells and Acini by Collagenase Digestion, Pancreapedia , , 2010
          大鼠大鼠, 成年的IsletsCollagenase Type 1: 450 u/mlLeibowitz L-15Williams, 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IsletsCollagenase Type 4: see referenceRPMI 1640Getty-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IsletsCollagenase Type 1: 450 u/mlRPMI 1640MacGregor, 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 isletsCollagenase: 0.75%RPMI 1640Tian 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 isletCollagenase Type 4: 0.2%HBSSHaefliger 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 aciniCLSPA: 30 u/mlM199Blinman 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AncinarCLSPA: see referenceDMEMJi 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IsletsCollagenase: 126 - 196 u/mlHBSSVerspohl, 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IsletsCollagenase: 0.5 - 0.9%HBSSTakaki, 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DuctPapain: 25 u/mlDMEM /F-12Githens, 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AcinarHyaluronidase: 462 u/mlHam's F12Hirschi, 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 acinarTrypsin: 0.001%F12 mediumYeh, 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 acinarTrypsin: 0.02%Solution BFoskett, 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IsletsCollagenase: 0.1% - 0.2%HBSSOhzato, 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 ductsPapain: 25 u/mlDMEM/Ham's F-12Githens, 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AcinarSoybean Trypsin Inhibitor: 0.01%HEPESMenozzi, 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, parotidHyaluronidase: 0.015%Earle's MEMMelvin, 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AcinarTrypsin: 0.01%HBSS, CMFOliver, 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EpithelialTrypsin: 0.1%Ham's F-12/HBSSTsao, 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IsletsCollagenase Type 4: 0.2%HBSSGotoh, 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 glandHyaluronidase: 0.1 %HBSS, CFQuissell, 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IsletsCollagenase Type 1: 0.3%Hank's solutionSutton, 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AcinarHyaluronidase: 0.1%HBSSBrannon, 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DuctTrypsin: 0.01%HBSSGithens, 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ExocrineHyaluronidase: 0.9%Kreb's RingerSchulz, 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IsletsCollagenase Type 4: 1%Medium 199Katada, 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 AcinarHyaluronidase: 0.18%Kreb's Henseleit bicarbonate bufferWilliams, 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IsletsCollagenase: 0.2%Kreb's Ringer bicarbonate bufferWolters, 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IsletsCollagenase Type 4: 0.5%HBSSShibata, 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ExocrineHyaluronidase: 0.15%KrebsKondo, 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IsletsCollagenase Type 4: 0.63%EBSSBraaten, 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, parotidTrypsin: 0.01%HBSS CMFMangos, 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IsletsTrypsin: 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IsletsCollagenase: 0.5%Hanks solutionLacy, 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IsletsCollagenase: 0.5%HBSSBallinger, 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IsletsCollagenase Type 4: 1.0% - 1.2%HBSSLacy, 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 hepatomaTrypsin: 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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