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Worthington:肾上腺组织分离指南

发布者:艾美捷科技    发布时间:2022-05-30     
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组织分离/原代细胞分离,是酶在组织培养研究和细胞生物学研究中的主要应用。尽管多年来在这些应用中广泛使用酶,但它们在解离和收获中的作用机制尚不清楚。


tissue-worthington.png研究人员在科学文献中搜索有关理想酶和组织解离的最佳条件的信息时,经常会遇到相互矛盾的数据。大部分变化源于细胞外基质的复杂性和动态性,以及在细胞分离应用中使用相对粗糙、不确定的酶制剂。

 

本指南总结了Worthington如何使用这些酶来完成对应肾上腺组织的解离,以及原代细胞采集的常规操作。 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

 

肾上腺 
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嗜铬细胞 / ChromaffinCollagenase Type 1: 0.1-0.2%;
Deoxyribonuclease I: 0.003-0.015%;
Hyaluronidase: 0.015%
DMEM/F12Dominguez, N., Rodri茆uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012
Bovine, 6 month嗜铬细胞 / ChromaffinCollagenase Type 1: 0.125%Locke's solutionMoustafa T, Girod S, Tortosa F, Li R, Sol JC, Rodriguez F, Bastide R, Lazorthes Y, Sallerin B: Viability and functionality of bovine chromaffin cells encapsulated into alginate-PLL microcapsules with a liquefied inner core, Cell Transplant 15, 121-33, 2006
Bovine嗜铬细胞 / Chromaffin cellsCollagenase: 0.1%;
Deoxyribonuclease I: 30 u/ml
DMEMHahm, S., Chen, Y., Vinson, C. and Eiden, L.: A Calcium-Initiated Signaling Pathway Propagated Through Calcineurin and cAMP Response Element-Binding Protein Activates Proenkephalin Gene Transcription after Depolarization., Mol Pharmacol 64, 1503, 2003
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.2%Locke's solutionOrtega, J., Sagen, J., and Pappas, G.: Short-term Immunosuppression Enhances Long-term Survival of Bovine Chromaffin Cell Xenografts in Rat CNS, Cell Transplant 1, 33, 1992
Bovine嗜铬细胞 / ChromaffinDeoxyribonuclease I: 30 u/mgHEPESZhu, J., Li, W., Toews, M., and Hexum, T.: Neuropeptide Y Inhibits Forskolin-Stimulated Adenylate Cyclase in Bovine Adrenal Chromaffin Cells via a Pertussis Toxin-Sensitive Process, J Pharmacol Exp Ther 263 (3), 1479, 1992
Bovine (also rat)心脏,肾上腺嗜铬,副神经元 / Heart,Adrenal chromaffin,ParaneuronsTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and Bovine Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
Bovine嗜铬细胞 / ChromaffinCollagenase Type 1: 0.25 %DMEMDahmer, M., Hart, P., and Perlman, R.: Studies on the Effect of Insulin-Like Growth Factor-I on Catecholamine Secretion from Chromaffin Cells, J Neurochem 54 (3), 931, 1990
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.2%;
Deoxyribonuclease I: 0.015%
Locke's solutionHiggins, L., and Berg, D.: Immunological Identification of a Nicotinic Acetylcholine Receptor on Bovine Chromaffin Cells, J Neurosci 7 (6), 1792, 1987
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.05%Locke's solution, CMFAunis, D., Rotllan, P., and Miras-Portugal, M.: Incorporation of Adenosine into Nucleotides of Chromaffin Cells in Culture, Neurochem Int 7, 89, 1985
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.15%Kreb's, CMFAlmazan, G., Aunis, D., Garcia, A., Montiel, C., Nicolas, G., and Sanchez-Garcia, P.: Effects Of CLS on the Release of Noradrenaline From Chromaffin Cells, Br J Biomed Sci 81, 599, 1984
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.1%(see reference)Pollard, H., Pazoles, C., Creutz, C., Scott, J., Zinder, O., and Hotchkiss, A.: An Osmotic Mechanism For Exocytosis From Dissociated Chromaffin Cells, J Biol Chem 259, 1114, 1984
Bovine嗜铬细胞 / ChromaffinCollagenase Type 1: 0.05%CF Kreb'sCena, V., Garcia, A., Montiel, C., and Sanchez-Garcia, P.: Uptake of [3H]-nicotine and [3H]-noradrenaline by Cultured Chromaffin Cells, Br J Pharmacol 81, 119, 1984
Bovine嗜铬细胞 / ChromaffinCollagenase Type 1: 0.025%HBSS, modifiedWaymire, J., Bennett, W., Boehme, R., Hankins, L., Gilmer-Waymire, K., and Haycock, J.: Bovine Adrenal Chromaffin Cells: High-Yield Purification and Viability in Suspension Culture, J Neurosci Methods 7, 329, 1983
Bovine髓质 / MedullaHyaluronidase: 0.2%Saline w/BSA 0.5%Knight, D. and Baker, P.: Stimulus-Secretion Coupling in Isolated Bovine Adrenal Medullary Cells, Q J Exp Physiol 68, 123, 1983
Bovine髓质 / MedullaCollagenase: 0.2%Krebs-Ringer bicarbonate buffer, CMFGreenberg, A. and Zinder, O.: alpha- and beta-Receptor Control of Catecholamine Secretion from Isolated Adrenal Medulla Cells, Cell Tissue Res 226, 655, 1982
Bovine嗜铬细胞 / ChromaffinDeoxyribonuclease I: 15 痢/mlMedium A (see reference)Wilson, S.P., and Viveros, O.H.: Primary Culture of Adrenal Medullary Chromaffin Cells in a Chemically Defined Medium, Exp Cell Res 133, 159, 1981
Bovine髓质 / MedullaProtease: 0.2%SalineBaker, P., and Knight., D: Calcium Control of Exocytosis and Endocytosis in Bovine Adrenal Medullary Cells, Phil Trans R Soc Lond 296, 83, 1981
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.05%Locke's solution, CMFTrifaro, J.M., and Lee, R.W.: Morphological Characteristics and Stimulus-Secretion Coupling in Bovine Adrenal Chromaffin Cell Cultures, Neuroscience 5, 1533, 1980
Bovine, adult (also rat, Hanover-Wistar, young; guinea-pig, newborn)嗜铬细胞 / ChromaffinCollagenase: 0.5%HBSSUnsicker, K., Rieffert, B., and Ziegler, W.: Effects of Cell Culture Conditions, Nerve Growth Factor, Dexamethasone, and Cyclic AMP on Adrenal Chromaffin Cells In Vitro, Adv Biochem Psychopharmacol 255, 51, 1980
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.25%F-12 mediumKumakura, K., Karoum, F., Guidotti, A., and Costa, E.: Modulation of Nicotinic Receptors by Opiate Receptor Agonists in Cultured Adrenal Chromaffin Cells, Nature 283, 489, 1980
Bovine髓质 / MedullaCollagenase: 0.05%Locke's solution, CFKilpatrick, D., Ledbetter, F., Carson, K., Kirshner, A., Slepetis, R., and Kirshner, N.: Stability of Bovine Adrenal Medulla Cells in Culture, J Neurochem 35 (3), 679, 1980
Bovine, adult髓质 / MedullaCollagenase Type 1: 0.5%HBSSUnsicker, K., and Griesser, G.: Establishment, Characterization and Fibre Outgrowth of Isolated Bovine Adrenal Medullary Cells in Long-Term Cultures, Neuroscience 5, 1445, 1980
Bovine嗜铬细胞 / ChromaffinCollagenase: 0.05%DMEMAunis, D., Guerold, B., Bader, M-F., and Cieselski-Treska, J.: Immunocytochemical and Biochemical Demonstration of Contractile Proteins in Chromaffin Cells, Neuroscience 5, 2261, 1980
Bovine髓质 / MedullaryCollagenase Type 2: 0.2%HEPESHersey, R., and DiStefano, V.: Control of Phenylethanolamine N-Methyltransferase by Glucocorticoids in Cultured Bovine Adrenal Medullary Cells, J Pharmacol Exp Ther 209 (1), 147, 1979
Bovine包皮 / ForeskinCollagenase: 0.5%Dulbecco's MEM w/10% calf serumFolkman, J., Haudenschild, C. C., and Zetter, B. R.: Long-term Culture of Capillary Endothlial Cells, Proc Natl Acad Sci U S A 76, 5217, 1979
Bovine髓质 / MedullaCollagenase Type 1: 0.05%Kreb's, CFFenwick, E., Fajdiga, P., Howe, N., and Livett, B.: Functional and Morphological Characterization of Isolated Bovine Adrenal Medullary Cell, J Cell Biol 76, 12, 1978
Bovine嗜铬细胞 / ChromaffinHyaluronidase: 0.2%HEPES, CFBrooks, J.C.: The Isolated Bovine Adrenomedullary Chromaffin Cell: A Model of Neuronal Excitation-Secretion, Endocrinology 101, 1369, 1977
Bovine, male嗜铬细胞 / ChromaffinCollagenase Type 1: 0.2%Kreb's Ringer bicarbonate buffer, CFSchneider, A., Herz, R., and Rosenheck, K.: Stimulus-Secretion Coupling in Chromaffin Isolated From Adrenal Medulla, Proc Natl Acad Sci U S A 74, 5036, 1977
豚鼠Guinea-pig, 500-700 g嗜铬,髓质 / Chromaffin,MedullaCollagenase:BSS (see reference)Role, L.W., Leeman, S.E., and Perlman, R.L.: Somatostain and Substance P Inhibit Catecholamine Secretion from Isolated Cells of Guinea-pig Adrenal Medulla, Neurochem Int 6, 1813, 1981
豚鼠Guinea-pig (also rat, Hanover-Wistar, young; newborn; cattle)嗜铬细胞 / ChromaffinCollagenase: 0.5%HBSSUnsicker, K., Rieffert, B., and Ziegler, W.: Effects of Cell Culture Conditions, Nerve Growth Factor, Dexamethasone, and Cyclic AMP on Adrenal Chromaffin Cells In Vitro, Adv Biochem Psychopharmacol 255, 51, 1980
豚鼠Guinea-pig肾上腺,嗜铬 / Adrenal,ChromaffinCollagenase: 0.05%-0.20%Kreb's-Ringer bicarb glucose buffer, CFHochman, J., and Perlman, R.L.: Catecholamine Secretion by Isolated Adrenal Cells, Biochim Biophys Acta 421, 168, 1976
仓鼠Hamster (Mesocricetus auratus) 100-150 g肾上腺,嗜铬 / Adrenal,ChromaffinHyaluronidase: 0.20%Kreb's Ringer bicarbonate bufferLiang, B.T., and Perlman, R.L.: Catecholamine Secretion by Hamster Adrenal Cells, J Neurochem 32, 927, 1979
Human肾上腺髓质祖细胞 / Adrenal medulla progenitorCollagenase: 0.2%DMEM/F-12Santana, M., Chung, K., Vukicevic, V., Rosmaninho-Salgado, J., Kanczkowski, W., Cortez, V., Hackmann, K., Bastos, C., Mota, A., Schrock, E., Bornstein, S., Cavadas, C. and Ehrhart-Bornstein, M.: Isolation, Characterization, and Differentiation of Progenitor Cells from Human Adult Adrenal Medulla., Stem Cells Transl Med 1, 783, 2012
Human嗜铬细胞 / Chromaffin cellsCollagenase: 0.2%Locke's solutionJeon, Y., Baek, W., Chung, S., Shin, N., Kim, H., and Lee, S.: Cultured Human Chromaffin Cells Grafted in Spinal Subarachnoid Space Relieves Allodynia in a Pain Rat Model., Korean J Anesthesiol 60, 357, 2011
Human肾上腺皮质 / AdrenocorticalCollagenase Type 1: 0.2%;
Deoxyribonuclease I: 0.01%
Krebs RingerCaroccia, B., Fassina, A., Seccia, T., Recarti, C., Petrelli, L., Belloni, A., Pelizzo, M. and Rossi, G.: Isolation of Human Adrenocortical Aldosterone-Producing Cells by a Novel Immunomagnetic Beads Method., Endocrinology 151, 1375, 2010
Human, adult嗜铬细胞 / ChromaffinTrypsin: 0.25%Eagle's MEMTischler, A., DeLellis, R., Bailes, B., Nunnemacher, G.,Carabba, V., and Wolfe, H.: Nerve Growth Factor-Induced Neurite Outgrowth from Normal Human Chromaffin Cells, Lab Invest 43, 399, 1980
Human, adult and child包皮 / ForeskinCollagenase: 0.5%Dulbecco's MEM w/10% calf serumFolkman, J., Haudenschild, C. C., and Zetter, B. R.: Long-term Culture of Capillary Endothlial Cells, Proc Natl Acad Sci U S A 76, 5217, 1979
小鼠Mouse嗜铬细胞 / ChromaffinCollagenase Type 1: 0.1-0.2%;
Deoxyribonuclease I: 0.003-0.015%;
Hyaluronidase: 0.015%
DMEM/F12Dominguez, N., Rodri茆uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012


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