組織分離/原代細胞分離，是酶在組織培養研究和細胞生物學研究中的主要應用。盡管多年來在這些應用中廣泛使用酶，但它們在解離和收獲中的作用機制尚不清楚。
研究人員在科學文獻中搜索有關理想酶和組織解離的最佳條件的信息時，經常會遇到相互矛盾的數據。大部分變化源于細胞外基質的復雜性和動態性，以及在細胞分離應用中使用相對粗糙、不確定的酶制劑。
本指南總結了Worthington如何使用這些酶來完成對應腎上腺組織的解離，以及原代細胞采集的常規操作。 本技術指南描述了標準實驗室程序； 為建立細胞分離方案提供了一種合乎邏輯的實驗方法； 并列出了許多組織特異性參考資料，以幫助開發有效的方法。更多其他組織的解離指南，點擊查看：各組織分離指南。
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| 種屬 | 種屬詳情 | 細胞名稱 | 酶組合推薦，含推薦濃度 | 培養基 | 參考文獻 |
| 牛 | Bovine | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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) | 心臟，腎上腺嗜鉻，副神經元 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 髓質 / 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 | 髓質 / 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 | 嗜鉻細胞 / 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 | 髓質 / 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 | 嗜鉻細胞 / 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) | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 髓質 / 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 | 髓質 / 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 | 嗜鉻細胞 / 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 | 髓質 / 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 | 髓質 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 | 嗜鉻，髓質 / 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) | 嗜鉻細胞 / 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 | 腎上腺，嗜鉻 / 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 |
| 倉鼠 | Hamster (Mesocricetus auratus) 100-150 g | 腎上腺，嗜鉻 / 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 | 腎上腺髓質祖細胞 / 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 | 嗜鉻細胞 / 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 | 腎上腺皮質 / 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 | 嗜鉻細胞 / 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 | 嗜鉻細胞 / 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 |
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