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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
Bovine导管上皮 / 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
Bovine (Bos taurus)导管 / 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
Bovine血小板 / PlateletsTrypsin:(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 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/狗Canine, adult mongrel, 15-25 kg胰岛 / IsletsCollagenase Type 4: 600-1100 u/ml;
Deoxyribonuclease I: 10 ug/ml
RPMI 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
Fish, Osphronemus gourami, 3-month-old, 0.5 mg胰岛 / 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Guinea-pig腺泡 / 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Guinea-pig, Hartley, albino, male, 350-400 g腺泡 / 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Guinea-pig外分泌 / 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
仓鼠Hamster胰岛 / 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
Human胰岛 / IsletsCollagenase Type 1: 1,600u/isolation;
Neutral Protease: 200 u/isolation;
Deoxyribonuclease I: 12,000 u/isolation
RPMIBrissova, 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
Human腺泡 / AcinarCLSPA: 200 u/ml;
Soybean Trypsin Inhibitor: 0.01%
See ReferenceCane, M., Sutton, R. and Criddle, D.: Isolation of Human Pancreatic Acinar Cells From Rat and Human Pancreas, Pancreapedia 20, 1, 2011
Human胰腺癌干细胞 / 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
Human, 25-55 yrs old胰岛 / 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, 615, 1995
Human胰岛 / IsletsTrypsin: Trypsin: 0.125-0.25%;
Collagenase: 0.2%;
Deoxyribonuclease I: 200 u/ml
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
Human, adult organ donors胰岛 / 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
Human胰岛 / 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
Human胰岛 / 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
Human, infant, age 1 day-1 year胰岛 / 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
Monkey, 3-5 Kg胰岛 / 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Mouse胰岛 / IsletsCollagenase Type 4: 0.1%HBSSLow, 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 / 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Mouse, 8-12 week胰腺类器官 / Pancreas organoidCollagenase: 0.012%;
Neutral Protease: 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
小鼠Mouse, embryonic胰祖细胞 / 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Mouse胰岛 / 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Mouse胰岛 / IsletCollagenase Type 4: 0.2%RPMI-1640Ding, 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 / 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., Cell 142, 787, 2010
小鼠Mouse, 6-8 month胰岛 / 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Mouse, adult, male安西纳尔 / AncinarCLSPA: see reference;
Soybean Trypsin Inhibitor: 0.001%
DMEMJi, 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 / 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Mouse胰岛 / 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Mouse胰岛 / 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Mouse胰岛 / 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Mouse, 6-10 month胰导管 / Pancreatic ductalCLSPA: 50 u/ml;
Hyaluronidase: 400 u/ml;
Soybean Trypsin Inhibitor: 0.02%
DMEMWang, 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 / 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Mouse, 11-12 week胰岛 / 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


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