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Worthington組織消化指南
引言
組織消化酶主要用于組織培養(yǎng)和細(xì)胞生物學(xué)研究的原代細(xì)胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年，但是對(duì)其組織消化和細(xì)胞收集的作用機(jī)制卻沒(méi)有全面的理解。 由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機(jī)選擇或基于過(guò)去的經(jīng)驗(yàn)，而不是在*理解消化原理的基礎(chǔ)上選擇的消化方案，以及如何根據(jù)具體實(shí)驗(yàn)?zāi)康男拚桨敢垣@得更好的實(shí)驗(yàn)結(jié)果。
細(xì)胞分離程序的目的是zui大程度的獲得功能活性的單個(gè)細(xì)胞，而影響結(jié)果的原因有很多，包括（不限于）以下：
1、組織類(lèi)型
2、物種來(lái)源
3、動(dòng)物年齡
4、遺傳改造情況（如knockouts）
5、所用的消化培養(yǎng)液
6、所用的酶
7、消化酶中的雜質(zhì)
8、所用消化酶的工作濃度
9、溫度
10、孵育時(shí)間
通常前四個(gè)原因沒(méi)辦法選擇，而剩下的原因一般由經(jīng)驗(yàn)決定以獲取zui適實(shí)驗(yàn)結(jié)果。
通過(guò)查閱文獻(xiàn)來(lái)選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實(shí)驗(yàn)數(shù)據(jù)搞得一頭霧水。這些巨大差異來(lái)源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性，以及過(guò)去常用的粗制、未提純的組織分離酶。另外，組織胞外基質(zhì)由各種蛋白、糖蛋白、脂類(lèi)和糖脂組成，這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育（年齡）之間存在明顯的差異，這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶，如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。
本指南總結(jié)了目前常用組織消化酶的zui適消化和細(xì)胞收集知識(shí)和方法，描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟， 以便于制定一個(gè)*的細(xì)胞分離方案，同時(shí)列舉了一些相關(guān)的參考文獻(xiàn)。
下表列出多種組織的適宜消化酶（含參考文獻(xiàn)，以組織類(lèi)型和物種分類(lèi)，詳見(jiàn)鏈接）
脂肪組織 | 腎上腺組織 | 骨組織 | 腦組織 |
軟骨組織 | 結(jié)腸組織 | 內(nèi)皮組織 | 上皮組織 |
眼 | 心臟 | 腸 | 腎 |
肝 | 肺 | 淋巴結(jié) | 乳腺 |
其它組織 | 其他組織 | 肌組織 | 神經(jīng)組織 |
胰腺 | 腮腺 | 垂體 | 前列腺 |
生殖 | 動(dòng)物鱗片 | 皮膚 | 脾臟 |
干細(xì)胞 | 胸腺 | 甲狀腺/甲狀旁腺 | 扁桃體 |
腫瘤 |
|
|
|
注：使用Worthington公司消化酶的文獻(xiàn)不限于上表鏈接中所列，一般而言，Worthington公司的組織消化酶可用于大多組織類(lèi)型的消化。
腎上腺組織分離手冊(cè)
| 物種 | 詳細(xì)來(lái)源 | 細(xì)胞名稱 | 消化用酶 | 培養(yǎng)基 | 參考文獻(xiàn) |
| 牛 | 牛 | Chromaffin | Collagenase Type 1: 0.1-0.2% | DMEM/F12 | Dominguez, N., Rodri璯uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012 |
| Deoxyribonuclease I: 0.003-0.015% | |||||
| Hyaluronidase: 0.015% | |||||
| 牛 | 牛, 6 個(gè)月 | 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 b綿羊 chromaffin cells encapsulated into alginate-PLL microcapsules with a liquefied inner core, Cell Transplant 15, 121-33, 2006 |
| 牛 | 牛 | Chromaffin cells | Collagenase: 0.1% | 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 |
| Deoxyribonuclease I: 30 u/ml | |||||
| 牛 | 牛 | Chromaffin | Collagenase: 0.2% | Locke's solution | Ortega, J., Sagen, J., and Pappas, G.: Short-term Immunosuppression Enhances Long-term Survival of B綿羊 Chromaffin Cell Xenografts in Rat CNS,Cell Transplant 1, 33, 1992 |
| 牛 | 牛 | Chromaffin | Deoxyribonuclease I: 30 u/mg | HEPES | Zhu, J., Li, W., Toews, M., and Hexum, T.: Neuropeptide Y Inhibits Forskolin-Stimulated Adenylate Cyclase in B綿羊 Adrenal Chromaffin Cells via a Pertussis Toxin-Sensitive Process, J Pharmacol Exp Ther 263 (3), 1479, 1992 |
| 牛 | 牛 | Heart | 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 B綿羊 Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990 |
| (also 大鼠) | Adrenal chromaffin | ||||
| Paraneurons | |||||
| 牛 | 牛 | 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 | 牛 | Chromaffin | Collagenase: 0.2% | Locke's solution | Higgins, L., and Berg, D.: Immunological Identification of a Nicotinic Acetylcholine Receptor on B綿羊 Chromaffin Cells, J Neurosci 7 (6), 1792, 1987 |
| Deoxyribonuclease I: 0.015% | |||||
| 牛 | 牛 | 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 | 牛 | 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 | 牛 | 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 | 牛 | 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 | 牛 | Chromaffin | Collagenase Type 1: 0.025% | HBSS, modified | Waymire, J., Bennett, W., Boehme, R., Hankins, L., Gilmer-Waymire, K., and Haycock, J.: B綿羊 Adrenal Chromaffin Cells: High-Yield Purification and Viability in Suspension Culture, J Neurosci Methods 7, 329, 1983 |
| 牛 | 牛 | Medulla | Hyaluronidase: 0.2% | Saline w/BSA 0.5% | Knight, D. and Baker, P.: Stimulus-Secretion Coupling in Isolated B綿羊 Adrenal Medullary Cells, Q J Exp Physiol 68, 123, 1983 |
| 牛 | 牛 | 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 Res226, 655, 1982 |
| 牛 | 牛 | Chromaffin | Deoxyribonuclease I: 15 礸/ml | Medium A | 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 |
| (see reference) | |||||
| 牛 | 牛 | Medulla | Protease: 0.2% | Saline | Baker, P., and Knight., D: Calcium Control of Exocytosis and Endocytosis in B綿羊 Adrenal Medullary Cells, Phil Trans R Soc Lond 296, 83, 1981 |
| 牛 | 牛 | Chromaffin | Collagenase: 0.05% | Locke's solution, CMF | Trifaro, J.M., and Lee, R.W.: Morphological Characteristics and Stimulus-Secretion Coupling in B綿羊 Adrenal Chromaffin Cell Cultures, Neuroscience 5, 1533, 1980 |
| 牛 | 牛, 成年的 | 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 |
| (還有大鼠, 加拿大-威斯塔, 年幼的; 豚鼠, 初生) | |||||
| 牛 | 牛 | 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 | 牛 | Medulla | Collagenase: 0.05% | Locke's solution, CF | Kilpatrick, D., Ledbetter, F., Carson, K., Kirshner, A., Slepetis, R., and Kirshner, N.: Stability of B綿羊 Adrenal Medulla Cells in Culture, J Neurochem 35 (3), 679, 1980 |
| 牛 | 牛, 成年的 | Medulla | Collagenase Type 1: 0.5% | HBSS | Unsicker, K., and Griesser, G.: Establishment, Characterization and Fibre Outgrowth of Isolated B綿羊 Adrenal Medullary Cells in Long-Term Cultures,Neuroscience 5, 1445, 1980 |
| 牛 | 牛 | 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 | 牛 | Medullary | Collagenase Type 2: 0.2% | HEPES | Hersey, R., and DiStefano, V.: Control of Phenylethanolamine N-Methyltransferase by Glucocorticoids in Cultured B綿羊 Adrenal Medullary Cells, J Pharmacol Exp Ther 209 (1), 147, 1979 |
| 牛 | 牛 | 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 | 牛 | Medulla | Collagenase Type 1: 0.05% | Kreb's, CF | Fenwick, E., Fajdiga, P., Howe, N., and Livett, B.: Functional and Morphological Characterization of Isolated B綿羊 Adrenal Medullary Cell, J Cell Biol 76, 12, 1978 |
| 牛 | 牛 | Chromaffin | Hyaluronidase: 0.2% | HEPES, CF | Brooks, J.C.: The Isolated B綿羊 Adrenomedullary Chromaffin Cell: A Model of Neuronal Excitation-Secretion, Endocrinology 101, 1369, 1977 |
| 牛 | 牛, 雄性 | 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 | 豚鼠, 500-700克 | Chromaffin | Collagenase: | BSS | Role, L.W., Leeman, S.E., and Perlman, R.L.: Somatostain and Substance P Inhibit Catecholamine Secretion from Isolated Cells of 豚鼠 Adrenal Medulla, Neurochem Int 6, 1813, 1981 |
| Medulla | (see reference) | ||||
| 豚鼠 | 豚鼠 | 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 |
| (還有大鼠,漢諾威-威斯塔, 年幼的; 初生; 牛) | |||||
| 豚鼠 | 豚鼠 | Adrenal | 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 |
| Chromaffin | |||||
| 倉(cāng)鼠 | 倉(cāng)鼠 (金倉(cāng)鼠) 100-150 克 | Adrenal | Hyaluronidase: 0.20% | Kreb's Ringer bicarbonate buffer | Liang, B.T., and Perlman, R.L.: Catecholamine Secretion by 倉(cāng)鼠 Adrenal Cells, J Neurochem 32, 927, 1979 |
| Chromaffin | |||||
| 人類(lèi) | 人類(lèi) | 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 人類(lèi) Adult Adrenal Medulla., Stem Cells Transl Med 1, 783, 2012 |
| 人類(lèi) | 人類(lèi) | Chromaffin cells | Collagenase: 0.2% | Locke's solution | Jeon, Y., Baek, W., Chung, S., Shin, N., Kim, H., and Lee, S.: Cultured 人類(lèi) Chromaffin Cells Grafted in Spinal Subarachnoid Space Relieves Allodynia in a Pain Rat Model., Korean J Anesthesiol 60, 357, 2011 |
| 人類(lèi) | 人類(lèi) | Adrenocortical | Collagenase Type 1: 0.2% | Krebs Ringer | Caroccia, B., Fassina, A., Seccia, T., Recarti, C., Petrelli, L., Belloni, A., Pelizzo, M. and Rossi, G.: Isolation of 人類(lèi) Adrenocortical Aldosterone-Producing Cells by a Novel Immunomagnetic Beads Method., Endocrinology 151, 1375, 2010 |
| Deoxyribonuclease I: 0.01% | |||||
| 人類(lèi) | 人類(lèi), 成年的 | 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 人類(lèi) Chromaffin Cells, Lab Invest 43, 399, 1980 |
| 人類(lèi) | 人類(lèi), 成年的和小孩 | 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 | 小鼠 | Chromaffin | Collagenase Type 1: 0.1-0.2% | DMEM/F12 | Dominguez, N., Rodri璯uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012 |
| Deoxyribonuclease I: 0.003-0.015% | |||||
| Hyaluronidase: 0.015% | |||||
| 小鼠 | 小鼠, 8-10 周 | Chromaffin | Papain: 40 u/ml | DMEM | Kolski-Andreaco, A., Cai, H., Currle, D., Chandy, K. and Chow, R.: 小鼠 Adrenal Chromaffin Cell Isolation., J Vis Exp 2, 129, 2007 |
| 小鼠 | 小鼠, 胚胎 | Chromafin cells | Papain: 20-25 u/ml | DMEM | Tian Jin-Hua, Wu Zheng-Xing, Unzicker Michael, Lu Li, Cai Qian, Li Cuiling, Schirra Claudia, Matti Ulf, Stevens David, Deng Chuxia, Rettig Jens, Sheng Zu-Hang: The role of Snapin in neurosecretion: snapin knock-out mice exhibit impaired calcium-dependent exocytosis of large dense-core vesicles in chromaffin cells, J Neurosci 25, 10546-55, 2005 |
| 綿羊 | 綿羊, 成年的和胚胎 | Adrenocortical | Collagenase Type 1: 0.4% | DMEM/Ham's F12 | Valego, N. and Rose, J.: A Specific CRH Antagonist Attenuates ACTH-Stimulated Cortisol Secretion in 綿羊 Adrenocortical Cells., Reprod Sci 17, 477, 2010 |
| 綿羊 | 綿羊, 胚胎 | Adrenocortical | Collagenase Type 1: 0.4% | DMEM/Ham's F12 | Valego, N., Su, Y., Carey, L., Young, S., Tatter, S., Wang, J. and Rose, J.:Hypothalamic-Pituitary Disconnection in Fetal Sheep Blocks the Peripartum Increases in Adrenal Responsiveness and Adrenal ACTH Receptor Expression.,Am J Physiol Regul Integr Comp Physiol 289, R410, 2005 |
| 綿羊 | 綿羊, 成年 | Chromaffin cells | Collagenase Type 2: 0.2% | Locke's solution | Keating, D., Rychkov, G., Adams, M., Holgert, H., McMillen, I.C. and Roberts, M.: Opioid Receptor Stimulation Suppresses the Adrenal Medulla Hypoxic Response in Sheep by Actions on Ca(2+) and K(+) Channels., J Physiol555, 489, 2004 |
| Deoxyribonuclease I: 100 u/ml | |||||
| 綿羊 | 綿羊, 3 年 | Anterior pituitary | Trypsin: 2.5% | DMEM | Canny B J, O'Farrell K A, Clarke I J, Tilbrook A J: The influence of sex and gonadectomy on the hypothalamo-pituitary-adrenal axis of the sheep, J Endocrinol 162, 215-25, 1999 |
| Deoxyribonuclease I: 0.004% | |||||
| 大鼠 | 大鼠 | Chromaffin | Collagenase Type 1: 0.1-0.2% | DMEM/F12 | Dominguez, N., Rodri璯uez, M., Machado, J. and Borges, R.: Preparation and Culture of Adrenal Chromaffin Cells., Methods Mol Biol 846, 223, 2012 |
| Deoxyribonuclease I: 0.003-0.015% | |||||
| Hyaluronidase: 0.015% | |||||
| 大鼠 | 大鼠 | Chromaffin | Collagenase Type 1: 0.26% | HBSS | Gilabert, J, Montalvo, G, and Artalejo A.: Rat Chromaffin Cells Primary Cultures: Standardization and Quality Assessment for Single-Cell Assays,Nat Protoc , 294, 2006 |
| Deoxyribonuclease I: 0.015% | |||||
| Hyaluronidase: 0.015% | |||||
| 大鼠 | 大鼠, SD | Chromaffin cells | Collagenase Type 1: 0.26% | HBSS | Gilabert, J: Necessary Conditions to Maintain Rat Adrenal Chromaffin Cells in Primary Culture,Cell Biology of the Chromaffin Cell,Borges, R. and Gandia, L., Instituto Teofilo Hernando, , 2004 |
| Deoxyribonuclease I: 0.015% | |||||
| Hyaluronidase: 0.015% | |||||
| 大鼠 | 大鼠, SD, 雄性 | Zona fasciculata/reticularis | Collagenase: 0.4% | Krebs-HEPES | Bruder Eric D, Ball Dennis L, Goodfriend Theodore L, Raff Hershel: An oxidized metabolite of linoleic acid stimulates corticosterone production by rat adrenal cells, Am J Physiol Regul Integr Comp Physiol 284, R1631-5, 2003 |
| 大鼠 | 大鼠, 威斯塔, 初生 | Chromaffin cells | Collagenase Type 1: 0.025% | DMEM | Zhang L, Casl A, Avila E, Drucker-Col鉚n R, Escobar A:Immunocytochemical, ultrastructural and neurochemical evidences on synaptogenesis and dopamine release of rat chromaffin cells co-cultured with striatal neurons, J Neuropathol Exp Neurol 59, 170-4, 2000 |
| Deoxyribonuclease I: 0.0015% | |||||
| 大鼠 | 大鼠, SD, 雄性 | ZG | Collagenase Type 1: 0.2% | Kreb's | Sayed, S., Whitehouse, B., and Jones, P.: Phosphoserine/Threonine Phosphatases in the Rat Adrenal Cortex: A Role in the Control of Steroidogenesis, J Endocrinol 154, 449, 1997 |
| ZFR | |||||
| 大鼠 | 大鼠, 費(fèi)希爾, 雄性, 10-16周 | Adrenocortical | Deoxyribonuclease I: 0.005% | BSS | Roskelley, C.D. and Auersperg, N.: Density Separation of Rat Adrenocortical Cells: Morphology, Steroidogenesis, and P-450scc Expression in Primary Culture,In Vitro Cell Dev Biol 26, 493, 1990 |
| 大鼠 | 大鼠, 雄性, 120-160 g, | Leydig | Collagenase Type 2: 0.03% (adrenal) | Krebs Ringer bicarbonate buffer | Ng, T. and Liu, W.: Toxic Effect of Heavy Metals on Cells Isolated from the Rat Adrenal and Testis, In Vitro Cell Dev Biol 26, 24, 1990 |
| 大鼠, SD, 雄性, 400-450 g | Adrenal | ||||
| 大鼠 | 大鼠, SD, 2-4 day old | Heart | 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 B綿羊 Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990 |
| (also 牛) | Adrenal chromaffin | ||||
| Paraneurons | |||||
| 大鼠 | 大鼠, Long-Evans, 雌性, 150-200 g | Glomerulosa | Collagenase: 0.2% | MEM-d-Val | Payet, N., Deziel, Y., and Lehoux, J.-G.: Vasopressin: A Potent Growth Factor in Adrenal Glomerulosa Cells in Culture, J Steroid Biochem 20, 449, 1984 |
| 大鼠 | 大鼠, 費(fèi)希爾, 雄性, 1-10月 | Adrenocortical | Deoxyribonuclease I: 0.005% | BSS | Leonard, R.K., Auersperg, N., and Parkes, C.O.: Ascorbic Acid Accumulation by Cultured Rat Adrenocortical Cells, In Vitro 19, 46, 1983 |
| 大鼠 | 大鼠, SD, 雄性, 400-450 克 | Decapular | Deoxyribonuclease I: 0.01% | Medium 199 | Li, C.H., Ng, T.B., and Cheng, C.H.K.: Melanotropins: Aldosterone- and Corticosterone-Stimulating Activity in Isolated Rat Adrenal Cells, Int J Pept Protein Res 19, 361, 1982 |
| Capsular | |||||
| Glomerulosa | |||||
| 大鼠 | 大鼠 | Chromaffin | Trypsin: 0.10% | Ham's F-12 w/HEPES | Englert, D.F.: An Optical Study of Isolated Rat Adrenal Chromaffin Cells, Exp Cell Res 125, 369, 1980 |
| 大鼠 | 大鼠, 漢諾威-威斯塔, 第二出生后的一周 (還有 豚鼠, 牛) | 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, Adv Biochem Psychopharmacol 25, 51, 1980 |
| 大鼠 | 大鼠 , SD, 雌性, 200 克 | Glomerulosa | Deoxyribonuclease I: 0.05% | Kreb's | Braley, L., Williams, G., and Bradwin, G.: The Effect of Unit Gravity Sedimentation on Adrenal Steroidogenesis by Isolated Rat Glomerulosa and Fasciculata Cells, Endocrinology 106 (1), 50, 1980 |
| 大鼠 | 大鼠 | 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 |
| 大鼠 | 大鼠, 漢諾威-威斯塔, 7-12 天 | Medullary | Trypsin: 0.125% | HBSS | Unsicker, K., Krisch, B., Otten, U., and Thoenen, H.: Nerve Growth Factor-Induced Fiber Outgrowth From Isolated Rat Adrenal Chromaffin Cells: Impairment by Glucocorticoids, Proc Natl Acad Sci U S A 75 (7), 3498, 1978 |
| 大鼠 | 大鼠, SD, 雄性 | Cortical | Trypsin: 0.25% | Kreb's Ringer bicarbonate buffer | Barofsky, A., Feinstein, M., and Halkerston, I.: Enzymatic and Mechanical Requirements for the Dissociation of Cortical Cells From Rat Adrenal Glands,Exp Cell Res 79, 263, 1973 |
| 大鼠 | 大鼠, Holtzman, 雄性, 180-250 g | Adrenal | Collagenase Type 1: 0.5% | Kreb's Ringer bicarbonate buffer | Kloppenborg, P., Island, D., Liddle, G., Michelakis, A., and Nicholson, W.: A Method of Preparing Adrenal Cell Suspensions and Its Applicability to the In Vitro Study of Adrenal Metabolism, Endocrinology 82, 1053, 1968 |
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