worthington 胰腺组织分离指南

发布者：艾美捷科技发布时间：2022-06-07

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原代细胞分离中，针对胰腺组织涉及到导管上皮，导管、血小板、颊神经节，胰岛和腺泡等等的解离通常会使用酶，如何选择合适的酶来完成对应胰腺组织的解离，以及原代细胞采集的常规操作， 艾美捷针对胰腺相关的组织分类解离分种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献，详细汇总指南： worthington胰腺组织分离指南

本技术指南描述了标准实验室程序；为建立细胞分离方案提供了一种合乎逻辑的实验方法；并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南。

乳腺
种属	种属详情	细胞名称	酶组合推荐，含推荐浓度	培养基	参考文献
牛	Bovine	导管上皮 / 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	Bovine (Bos taurus)	导管 / 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	Bovine	血小板 / Platelets	Trypsin:	(see reference)	Stiles, G., and Lefkowitz, R.: Hormone-Sensitive Adneylate Cyclase, J Biol Chem 257 (11), 6287, 1982
犬/狗	Canine	颊神经节， SLT 肌肉 / 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/狗	Canine, adult mongrel, 15-25 kg	胰岛 / Islets	Collagenase Type 4: 600-1100 u/ml； Deoxyribonuclease I: 10 ug/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
鱼	Fish, Osphronemus gourami, 3-month-old, 0.5 mg	胰岛 / 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	Guinea-pig	腺泡 / 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	Guinea-pig, Hartley, albino, male, 350-400 g	腺泡 / 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	Guinea-pig	外分泌 / 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
仓鼠	Hamster	胰岛 / 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	Human	胰岛 / Islets	Collagenase Type 1: 1,600u/isolation； Neutral Protease: 200 u/isolation； Deoxyribonuclease I: 12,000 u/isolation	RPMI	Brissova, M., Haliyur, R., Saunders, D., Shrestha, S., Dai, C., Blodgett, D., Bottino, R., Campbell-Thompson, M., Aramandla, R., Poffenberger, G., Lindner, J., Pan, F., von Herrath, M., Greiner, D. and Shultz, L.: α Cell Function and Gene Expression Are Compromised in Type 1 Diabetes., Cell Rep 22, 2667-2676, 2018
人	Human	腺泡 / Acinar	CLSPA: 200 u/ml； Soybean Trypsin Inhibitor: 0.01%	See Reference	Cane, M., Sutton, R. and Criddle, D.: Isolation of Human Pancreatic Acinar Cells From Rat and Human Pancreas, Pancreapedia 20, 1, 2011
人	Human	胰腺癌干细胞 / 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	Human, 25-55 yrs old	胰岛 / 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, 615, 1995
人	Human	胰岛 / Islets	Trypsin: Trypsin: 0.125-0.25%； Collagenase: 0.2%； Deoxyribonuclease I: 200 u/ml	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
人	Human, adult organ donors	胰岛 / 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	Human	胰岛 / 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	Human	胰岛 / 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	Human, infant, age 1 day-1 year	胰岛 / 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	Monkey, 3-5 Kg	胰岛 / 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	Mouse	胰岛 / Islets	Collagenase Type 4: 0.1%	HBSS	Low, L., Zavortink, M., Mitchell, J., Gan, W., Do, O., Schwiening, C., Gaisano, H. and Thorn, P.: Insulin Secretion from Beta Cells in Intact Mouse Islets is Targeted Towards the Vasculature, Diabetologia 57, 1655, 2014
小鼠	Mouse	胰岛 / 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	Mouse, 8-12 week	胰腺类器官 / Pancreas organoid	Collagenase: 0.012%； Neutral Protease: 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
小鼠	Mouse, embryonic	胰祖细胞 / 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	Mouse	胰岛 / 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	Mouse	胰岛 / Islet	Collagenase Type 4: 0.2%	RPMI-1640	Ding, G., Wang, F., Shu, J., Tian, S., Jiang, Y., Zhang, D., Wang, N., Luo, Q., Zhang, Y., Jin, F., Leung, P., Sheng, J. and Huang, H.: Transgenerational Glucose Intolerance with Igf2/H19 Epigenetic Alterations in Mouse Islet Induced by Intrauterine Hyperglycemia., Diabetes 61, 1133-42, 2012
小鼠	Mouse, 10 week	胰岛 / 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., Cell 142, 787, 2010
小鼠	Mouse, 6-8 month	胰岛 / 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	Mouse, adult, male	安西纳尔 / Ancinar	CLSPA: see reference； Soybean Trypsin Inhibitor: 0.001%	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
小鼠	Mouse, 3-4 week, 20-24 g	胰岛 / 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	Mouse	胰岛 / 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	Mouse	胰岛 / 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	Mouse	胰岛 / 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	Mouse, 6-10 month	胰导管 / Pancreatic ductal	CLSPA: 50 u/ml； Hyaluronidase: 400 u/ml； Soybean Trypsin Inhibitor: 0.02%	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
小鼠	Mouse, adult	腺泡 / 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	Mouse, 11-12 week	胰岛 / 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