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。

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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase Type 1: 0.1-0.2%； Deoxyribonuclease I: 0.003-0.015%； Hyaluronidase: 0.015%	DMEM/F12	Dominguez, N., Rodri茆uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012
牛	Bovine, 6 month	嗜铬细胞 / Chromaffin	Collagenase Type 1: 0.125%	Locke's solution	Moustafa 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
牛	Bovine	嗜铬细胞 / Chromaffin cells	Collagenase: 0.1%； Deoxyribonuclease I: 30 u/ml	DMEM	Hahm, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.2%	Locke's solution	Ortega, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Deoxyribonuclease I: 30 u/mg	HEPES	Zhu, 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
牛	Bovine (also rat)	心脏，肾上腺嗜铬，副神经元 / Heart，Adrenal chromaffin，Paraneurons	Trypsin: 0.06%	25mM HEPES buffered Locke's solution, CMF	Trifaro, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase Type 1: 0.25 %	DMEM	Dahmer, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.2%； Deoxyribonuclease I: 0.015%	Locke's solution	Higgins, L., and Berg, D.: Immunological Identification of a Nicotinic Acetylcholine Receptor on Bovine Chromaffin Cells, J Neurosci 7 (6), 1792, 1987
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.05%	Locke's solution, CMF	Aunis, D., Rotllan, P., and Miras-Portugal, M.: Incorporation of Adenosine into Nucleotides of Chromaffin Cells in Culture, Neurochem Int 7, 89, 1985
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.15%	Kreb's, CMF	Almazan, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase Type 1: 0.05%	CF Kreb's	Cena, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase Type 1: 0.025%	HBSS, modified	Waymire, 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
牛	Bovine	髓质 / Medulla	Hyaluronidase: 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
牛	Bovine	髓质 / Medulla	Collagenase: 0.2%	Krebs-Ringer bicarbonate buffer, CMF	Greenberg, A. and Zinder, O.: alpha- and beta-Receptor Control of Catecholamine Secretion from Isolated Adrenal Medulla Cells, Cell Tissue Res 226, 655, 1982
牛	Bovine	嗜铬细胞 / Chromaffin	Deoxyribonuclease I: 15 痢/ml	Medium 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
牛	Bovine	髓质 / Medulla	Protease: 0.2%	Saline	Baker, P., and Knight., D: Calcium Control of Exocytosis and Endocytosis in Bovine Adrenal Medullary Cells, Phil Trans R Soc Lond 296, 83, 1981
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.05%	Locke's solution, CMF	Trifaro, J.M., and Lee, R.W.: Morphological Characteristics and Stimulus-Secretion Coupling in Bovine Adrenal Chromaffin Cell Cultures, Neuroscience 5, 1533, 1980
牛	Bovine, adult (also rat, Hanover-Wistar, young; guinea-pig, newborn)	嗜铬细胞 / Chromaffin	Collagenase: 0.5%	HBSS	Unsicker, 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
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.25%	F-12 medium	Kumakura, 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
牛	Bovine	髓质 / Medulla	Collagenase: 0.05%	Locke's solution, CF	Kilpatrick, 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
牛	Bovine, adult	髓质 / Medulla	Collagenase Type 1: 0.5%	HBSS	Unsicker, K., and Griesser, G.: Establishment, Characterization and Fibre Outgrowth of Isolated Bovine Adrenal Medullary Cells in Long-Term Cultures, Neuroscience 5, 1445, 1980
牛	Bovine	嗜铬细胞 / Chromaffin	Collagenase: 0.05%	DMEM	Aunis, D., Guerold, B., Bader, M-F., and Cieselski-Treska, J.: Immunocytochemical and Biochemical Demonstration of Contractile Proteins in Chromaffin Cells, Neuroscience 5, 2261, 1980
牛	Bovine	髓质 / Medullary	Collagenase Type 2: 0.2%	HEPES	Hersey, 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
牛	Bovine	包皮 / Foreskin	Collagenase: 0.5%	Dulbecco's MEM w/10% calf serum	Folkman, 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
牛	Bovine	髓质 / Medulla	Collagenase Type 1: 0.05%	Kreb's, CF	Fenwick, E., Fajdiga, P., Howe, N., and Livett, B.: Functional and Morphological Characterization of Isolated Bovine Adrenal Medullary Cell, J Cell Biol 76, 12, 1978
牛	Bovine	嗜铬细胞 / Chromaffin	Hyaluronidase: 0.2%	HEPES, CF	Brooks, J.C.: The Isolated Bovine Adrenomedullary Chromaffin Cell: A Model of Neuronal Excitation-Secretion, Endocrinology 101, 1369, 1977
牛	Bovine, male	嗜铬细胞 / Chromaffin	Collagenase Type 1: 0.2%	Kreb's Ringer bicarbonate buffer, CF	Schneider, 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，Medulla	Collagenase:	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 / Chromaffin	Collagenase: 0.5%	HBSS	Unsicker, 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，Chromaffin	Collagenase: 0.05%-0.20%	Kreb's-Ringer bicarb glucose buffer, CF	Hochman, 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，Chromaffin	Hyaluronidase: 0.20%	Kreb's Ringer bicarbonate buffer	Liang, B.T., and Perlman, R.L.: Catecholamine Secretion by Hamster Adrenal Cells, J Neurochem 32, 927, 1979
人	Human	肾上腺髓质祖细胞 / Adrenal medulla progenitor	Collagenase: 0.2%	DMEM/F-12	Santana, 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
人	Human	嗜铬细胞 / Chromaffin cells	Collagenase: 0.2%	Locke's solution	Jeon, 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
人	Human	肾上腺皮质 / Adrenocortical	Collagenase Type 1: 0.2%； Deoxyribonuclease I: 0.01%	Krebs Ringer	Caroccia, 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
人	Human, adult	嗜铬细胞 / Chromaffin	Trypsin: 0.25%	Eagle's MEM	Tischler, 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
人	Human, adult and child	包皮 / Foreskin	Collagenase: 0.5%	Dulbecco's MEM w/10% calf serum	Folkman, 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 / Chromaffin	Collagenase Type 1: 0.1-0.2%； Deoxyribonuclease I: 0.003-0.015%； Hyaluronidase: 0.015%	DMEM/F12	Dominguez, N., Rodri茆uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012