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世界*實驗材料供應(yīng)商 Worthington Biochemical 正式授權(quán)上海起發(fā)為其中國代理， Worthington Biochemical 在一直是行業(yè)的*,一直為廣大科研客戶提供zui為的產(chǎn)品和服務(wù),上海起發(fā)一直秉承為中國科研客戶帶來的產(chǎn)品，的服務(wù)，簽約 Worthington Biochemical 就是為了給廣大科研客戶帶來更加完善的產(chǎn)品和服務(wù),您的滿意將是我們zui大的收獲
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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)和細胞生物學(xué)研究的原代細胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年，但是對其組織消化和細胞收集的作用機制卻沒有全面的理解。 由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機選擇或基于過去的經(jīng)驗，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)具體實驗?zāi)康男拚桨敢垣@得更好的實驗結(jié)果。
細胞分離程序的目的是zui大程度的獲得功能活性的單個細胞，而影響結(jié)果的原因有很多，包括（不限于）以下：
1、組織類型
2、物種來源
3、動物年齡
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、孵育時間
通常前四個原因沒辦法選擇，而剩下的原因一般由經(jīng)驗決定以獲取zui適實驗結(jié)果。
通過查閱文獻來選擇*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驗數(shù)據(jù)搞得一頭霧水。這些巨大差異來源于組織胞外基質(zhì)的復(fù)雜而動態(tài)的特性，以及過去常用的粗制、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適消化和細胞收集知識和方法，描述了標(biāo)準(zhǔn)的實驗步驟， 以便于制定一個*的細胞分離方案，同時列舉了一些相關(guān)的參考文獻。
下表列出多種組織的適宜消化酶（含參考文獻，以組織類型和物種分類，詳見鏈接）
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 | 動物鱗片 | 皮膚 | 脾臟 |
干細胞 | 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型的消化。
腦組織分離手冊
| 物種 | 詳細來源 | 細胞名稱 | 消化用酶 | 培養(yǎng)基 | 參考文獻 |
| 牛 | 牛 | Microvascular endothelial | Neutral Protease: 0.005% | Medium 199 | Kanda, T., Yoshino, H., Ariga, T., Yamawaki, M., and Yu, R.: Glycosphingolipid Antigens in Cultured Microvasular Bovine Brain Endothelial Cells: Sulfoglucuronosyl Paragloboside as a Target of Monoclonal IgM in Demyelinative Neuropathy, J Cell Biol 126 (1), 235, 1994 |
| 牛 | 牛 | Brain endothelial cells | Collagenase Type 2: 0.35% | DMEM | Wolburg H, Neuhaus J, Kniesel U, Krauss B, Schmid EM, Ocalan M, Farrell C, Risau W: Modulation of tight junction structure in blood-brain barrier endothelial cells. Effects of tissue culture, second messengers and cocultured astrocytes, J Cell Sci 107, 1347, 1994 |
| 牛 | 牛 | Endothelial | Neutral Protease: 0.125% | MEM | Miller, D., Audus, K., and Borchardt, R.: Application of Cultured Endothelial Cells of the Brain Microvasculature in the Study of the Blood-Brain Barrier, J Tiss Cul Meth 14, 217, 1992 |
| 牛 | 牛, 成年的 | Cerebral artery | Collagenase: 0.2% | HBSS | Machi, T., Kassell, N.F., and Scheld, W.M.: Isolation and Characterization of Endothelial Cells From Bovine Cerebral Arteries, In Vitro Cell Dev Biol 26, 291, 1990 |
| Endothelial | |||||
| 牛 | 牛 | Capillary endothelial | Collagenase: 0.1% | DMEM | Estrada, C., Bready, J., Berliner, J., and Cancilla, P.: Choline Uptake by Cerebral Capillary Endothelial Cells in Culture, J Neurochem 54, 1467, 1990 |
| 牛 | 牛 | Endothelial | Neutral Protease: 0.5% | MEM | Audus, K., and Borchardt, R.: Characterization of an In Vitro Blood-Brain Barrier Model System for Studying Drug Transport and Metabolism, Pharm Res 3 (2), 81, 1986 |
| 牛 | 牛 | Endothelial | Collagenase Type 2: 0.2% | Dulbecco's PBS | Goetz, I.., Warren, J., Estrada, C., Roberts, E., and Krause, D.: Long-Term C*tion of Arterial and Capillary Endothelium From 成年的 Bovine Brain, In Vitro Cell Dev Biol 21, 172, 1985 |
| Brain arteries | |||||
| 牛 | Calf | Oligodendroglia | Trypsin: 0.1% | (see reference) | Poduslo, S., Miller, K., and McKhann, G.: Metabolic Properties of Maintained Oligodendroglia Purified from Brain, J Biol Chem 253, 1592, 1978 |
| (also lamb) | Neural | ||||
| 豚鼠 | 豚鼠, 200-400 g | Neurons | Trypsin: 0.06-0.08% | PIPES saline | Kay, A.R., and Wong, R.K.S.: Isolation of Neurons Suitable for Patch-Clamping from 成年的 Mammalian Central Nervous Systems, J Neurosci Methods 16, 227, 1986 |
| 人類 | 人類 | Tumor | Collagenase Type 4: 0.1% | see reference | Sauvageot, C., Weatherbee, J., Kesari, S., Winters, S., Barnes, J., Dellagatta, J., Ramakrishna, N., Stiles, C., Kung, A., Kieran, M. and Wen, P.: Efficacy of the HSP90 Inhibitor 17-AAG in Human Glioma Cell Lines and Tumorigenic Glioma Stem Cells.,Neuro Oncol Vol. 11,, , 109, 2009 |
| Hyaluronidase: 0.07% | |||||
| Deoxyribonuclease I: 0.04% | |||||
| 人類 | 人類 | Microglia | Trypsin: 0.25% | DMEM/F12 | Klegeris Andis, McGeer PatrickL: Chymotrypsin-like proteases contribute to human monocytic THP-1 cell as well as human microglial neurotoxicity, Glia 51, 56-64, 2005 |
| Deoxyribonuclease I: .005% | |||||
| 人類 | 人類 | Microvessels | Collagenase Type 4: 0.1% | DMEM | Gerhart, D. Z., Broderius, M. A., and Drewes, L. R.: Cutlured Human and Canine Endothelial Cells from Brain Microvessels, Brain Res Bull 21, 785, 1988 |
| 人類 | 人類 | Neuronal | Deoxyribonuclease I: 10 礸/ml | Tris-HCl, 50 mM, CaCl2, 2 mM | Roher, A.E., Palmer, K.C., Chau, V., and Ball, M.J.: Isolation and Chemical Characterization of Alzheimer's Disease Paired Helical Filament Cytoskeletons:Differentiation from Amyloid Plaque Core Protein, J Cell Biol 107, 2703, 1988 |
| 人類 | 人類, 15-54 歲 | Microvessels | Collagenase: 0.1% | Serum-free modified Lewis medium | Vinters, H.V., Reave, S., Coslo, P., Girvin, J.P., and Moore, S.A.: Isolation and Culture of Cells Derived From Human Cerebral Microvessels, Cell Tissue Res 249, 657, 1987 |
| 昆蟲 | 果蠅 | Neurons | Papain: 20 u/ml | Saline | Gu, H. and O'Dowd, D.: Whole Cell Recordings from Brain of 成年的 Drosophila., J Vis Exp 6, 248, 2007 |
| 昆蟲 | 果蠅 | Neurons | Papain: 4 u/ml | DMEM | Sicaeros, B. Campusano, J. and O'Dowd, D.: Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae., J Vis Exp 4, 200, 2007 |
| 猴 | 猴, 獼猴 | Brain cells | Collagenase Type 2: 500 u/ml | HBSS | Marcondes MC, Burudi EM, Huitron-Resendiz S, Sanchez-Alavez M, Watry D, Zandonatti M, Henriksen SJ, Fox HS: Highly activated CD8(+) T cells in the brain correlate with early central nervous system dysfunction in simian immunodeficiency virus infection, J Immunol 167, 5429-38, 2001 |
| Deoxyribonuclease I: 28 u/ml | |||||
| 小鼠 | 小鼠 | Chloroid plexus | Collagenase Type 4: 400 u/ml | PBS | Baruch, K., Rosenzweig, N., Kertser, A., Deczkowska, A., Sharif, A., Spinrad, A., Tsitsou-Kampeli, A., Sarel, A., Cahalon, L. and Schwartz, M.: Breaking Immune Tolerance by Targeting Foxp3(+) Regulatory T Cells Mitigates Alzheimer's Disease Pathology., Nat Commun 6, 7967, 2015 |
| 小鼠 | 小鼠 | Brain-sequestered leukocytes | Collagenase Type 4: 0.05% | RPMI | Ryg-Cornejo, V., Ioannidis, L. and Hansen, D.: Isolation and Analysis of Brain-Sequestered Leukocytes from Plasmodium Berghei ANKA-Infected Mice., J Vis Exp71, e50112, 2013 |
| Deoxyribonuclease I: 2 u/ml | |||||
| 小鼠 | 小鼠 | Cerebral pericytes | Collagenase Type 2: 0.1% | DMEM | Shah. G., Price, T., Banks, W., Morofuji, Y., Kovac, A., Ercal, N., Sorenson, C., Shin, E., Sheibani, N.: Pharmacological Inhibition of Mitochondrial Carbonic Anhydrases Protects Mouse Cerebral Pericytes from High Glucose-Induced Oxidative Stress and Apoptosis., J Pharmacol Exp Ther 344, 637-45, 2013 |
| Deoxyribonuclease I: 30 u/ml | |||||
| 小鼠 | 小鼠, P13-15 | Glial | Papain: 20 u/ml | EBSS | Haseleu, J., Anlauf, E., Blaess, S., Endl, E. and Derouiche, A.: Studying Subcellular Detail in Fixed Astrocytes: Dissociation of Morphologically Intact Glial Cells (DIMIGs)., Front Cell Neurosci 7, 54, 2013 |
| Deoxyribonuclease I: 0.0005% | |||||
| 小鼠 | 小鼠, 成年的 | Neurons | PDS kit: with modifications | DMEM/F12 | Saxena, A., Wagatsuma, A., Noro, Y., Kuji, T., Asaka-Oba, A., Watahiki, A., Gurnot, C., Fagiolini, M., Hensch, T. and Carninci, P.: Trehalose-Enhanced Isolation of Neuronal Sub-Types from 成年的 Mouse Brain., Biotechniques 52, 381, 2012 |
| 小鼠 | 小鼠, 1-2 天 | Oligodendrocytes, dorsal root ganglia | Papain: 0.15% | DMEM | O'Meara, R., Ryan, S., Colognato, H. and Kothary, R.: Derivation of Enriched Oligodendrocyte Cultures and Oligodendrocyte/Neuron Myelinating Co-Cultures from Post-Natal Murine Tissues., J Vis Exp 54, 3324, 2011 |
| Deoxyribonuclease I: 0.006% | |||||
| 小鼠 | 小鼠, 6-12周 | Vascular smooth muscle cells | Papain: 0.05% | PBS | Chung, W., Farley, J., Swenson, A., Barnard, J., Hamilton, G., Chiposi, R. and Drummond, H.: Extracellular Acidosis Activates ASIC-like Channels in Freshly Isolated Cerebral Artery Smooth Muscle Cells., Am J Physiol Cell Physiol 298, C1198, 2010 |
| Collagenase Type 4: 0.15% | |||||
| Elastase: 0.05% | |||||
| 小鼠 | 小鼠, 4-6 天 | Neurons | PDS kit: with modifications | HBSS | Lee, H., Greene, L., Mason, C. and Manzini, M.: Isolation and Culture of Post-Natal Mouse Cerebellar Granule Neuron Progenitor Cells and Neurons., J Vis Exp23, 990, 2009 |
| 小鼠 | 小鼠, 產(chǎn)后的 | Astrocytes | Trypsin: 0.25% | HBSS | Sher, F., Rossler, R., Brouwer, N., Balasubramaniyan, V., Boddeke, E. and Copray, S.: Differentiation of Neural Stem Cells Into Oligodendrocytes: Involvement of the Polycomb Group Protein Ezh2., Stem Cells 26, 2875, 2008 |
| Deoxyribonuclease I: 1,000 u/ml | |||||
| 小鼠 | 小鼠, 初生的 | Neurons | Papain: 20 u/ml | Neurobasal | Fasano, C., Thibault, D. and Trudeau, L.: Culture of Postnatal Mesencephalic Dopamine Neurons on an Astrocyte Monolayer,Current Protocols in Neuroscience Vol. 44,, , 3.21.1, 2008 |
| 小鼠 | 小鼠, 胚胎和出生后 | Cortical neurons | Papain: 4-10 u/ml | Neurobasal | Hilgenberg, L. and Smith, M.: Preparation of Dissociated Mouse Cortical Neuron Cultures., J Vis Exp 10, 562, 2007 |
| 小鼠 | 小鼠 | Neurosperes | PDS kit: per instructions | DMEM/F12 | Klein C, Butt SJ, Machold RP, Johnson JE, and Fishell G.: Cerebellum- and forebrain-derived stem cells possess intrinsic regional character, Development132, 4497, 2005 |
| 小鼠 | 小鼠, 1 天 | Neural progenitor | PDS kit: per instructions | see reference | Seaberg, R., Smukler, S. and Van der Kooy, D.: Intrinsic Differences Distinguish Transiently Neurogenic Progenitors from Neural Stem Cells in the Early Postnatal Brain., Dev Biol 278, 71, 2005 |
| 小鼠 | 小鼠 | Granule cell precursors, pre-neoplastic and tumor cells | Papain: 10 u/ml | Neurobasal/B27 | Oliver, T., Read, T., Kessler, J., Mehmeti, A., Wells, J., Huynh, T., Lin, S. and Wechsler-Reya, R.: Loss of Patched and Disruption of Granule Cell Development in a Pre-Neoplastic Stage of Medulloblastoma., Development 132, 2425, 2005 |
| Deoxyribonuclease I: 250 u/ml | |||||
| 小鼠 | 小鼠 | Cortical neurons | PDS kit: per instructions | Neurobasal | Hernandez, F., Perez, M., Lucas, J., Mata, A., Bhat, R. and Avila, J.: Glycogen Synthase Kinase-3 plays a Crucial Role in Tau Exon 10 Splicing and Intranuclear Distribution of SC35. Implications for Alzheimer's Disease., J Biol Chem 279, 3801, 2004 |
| 小鼠 | 小鼠, 3 天 | Microglia | Papain: 90 u/ml | Eagle's MEM | Nishioku T, Hashimoto K, Yamashita K, Liou SY, Kagamiishi Y, Maegawa H, Katsube N, Peters C, von Figura K, Saftig P, Katunuma N, Yamamoto K, Nakanishi H: Involvement of cathepsin E in exogenous antigen processing in primary cultured murine microglia, J Biol Chem 277, 4816, 2002 |
| Deoxyribonuclease I: 2000 u/ml | |||||
| 小鼠 | 小鼠, SD, 8-12 周 | Microglia | Trypsin: 0.125% | RPMI-1640 | O'Donnell SL, Frederick TJ, Krady JK, Vannucci SJ, Wood TL: IGF-I and microglia/macrophage proliferation in the ischemic mouse brain, Glia 39, 85, 2002 |
| Collagenase Type 2: 0.01% | |||||
| Deoxyribonuclease I: .005% | |||||
| 小鼠 | 小鼠, 2-5天 | Postnal substantia nigra | PDS kit: per instructions | see reference | Smeyne Michelle, Smeyne RichardJ: Method for culturing postnatal substantia nigra as an in vitro model of experimental Parkinson's disease, Brain Res Brain Res Protoc 9, 105-11, 2002 |
| 小鼠 | 小鼠, 胚胎 | Cortical progenitors | PDS kit: per instructions | Serum free medium | Estivill-Torrus, G., Pearson, H., Van Heyningen, V., Price, D. and Rashbass, P.:Pax6 is Required to Regulate the Cell Cycle and the Rate of Progression from Symmetrical to Asymmetrical Division in Mammalian Cortical Progenitors.,Development 129, 455, 2002 |
| 小鼠 | 小鼠, 1 天 | Neurons and glia | PDS kit: per instructions | Neurobasal | Martin-Aparicio, E., Yamamoto, A., Hernandez, F., Hen, R., Avila, J. and Lucas, J.: Proteasomal-Dependent Aggregate Reversal and Absence of Cell Death in a Conditional Mouse Model of Huntington's Disease., J Neurosci 21, 8772, 2001 |
| 小鼠 | 小鼠 | Cerebellar granule cell precursors | Papain: 0.435% | EBSS | Miyazawa K, Himi T, Garcia V, Yamagishi H, Sato S, and Ishizaki Y.: A role for p27/Kip1 in the control of cerebellar granule cell precursor proliferation, J Neurosci 20, 5756, 2000 |
| Deoxyribonuclease I: 0.05% | |||||
| 小鼠 | 小鼠 | Astrocytes | Trypsin: 0.25% | DMEM | Lim, D, and Alvarez-Buylla, A.: Interaction Between Astrocytes and 成年的 Subventricular Zone Precursors Stimulates Neurogenesis, Proc Natl Acad Sci U S A 96, 7526, 1999 |
| 小鼠 | 小鼠, 初生的 | Hippocampal cells | Papain: 10 u/ml | DMKM | Jun K, Choi G, Yang SG, Choi KY, Kim H, Chan GC, Storm DR, Albert C, Mayr GW, Lee CJ, Shin HS: Enhanced hippocampal CA1 LTP but normal spatial learning in inositol 1,4,5-trisphosphate 3-kinase(A)-deficient mice, Learn Mem 5, 317-30, 1998 |
| 小鼠 | 小鼠, SWR or CF1, 1-3 個月 | Papillae, taste receptor | Pronase E: 0.15% | Carbonate-Phosphate buffer | Spielman, A., Mody, I., Brand, J., Whitney, G., MacDonald, J., and Salter, M.: A Method for Isolating and Patch-Clamping Single Mammalian Taste Receptor Cells,Brain Res 503, 326, 1989 |
| (see reference) | |||||
| 小鼠 | 小鼠, 0-30 天 | Neural | Trypsin NF 1:250: 50 0.25% | BSS | Shrier, B., Wilson, S., and Nirenberg, M.: Cultured Cell Systems and Methods for Neurobiology, Vol. 32,, , 765, 1974 |
| 綿羊 | 羔羊 | Oligodendroglia | Trypsin: 0.1% | (see reference) | Poduslo, S., Miller, K., and McKhann, G.: Metabolic Properties of Maintained Oligodendroglia Purified from Brain, J Biol Chem 253, 1592, 1978 |
| (還有 牛犢) | Neural | ||||
| 豬 | 小型豬, 尤卡坦半島 (尤卡坦半島野豬), 4-6 個月 | Microvascular | Collagenase: 0.1% | HBSS | Robinson, D.H., Kang,Y., Deschner, S.H., and Nielsen, T.B.: Morphologic Plasticity and Periodicity : Porcine Cerebral Microvascular Cells in Culture, In Vitro Cell Dev Biol 26, 169, 1990 |
| 大鼠 | 大鼠, SD, 19-21 天 | Suprachiasmatic nucleus neurons | Papain: 100 u/ml | MEM | Cao, R., Li, A., Cho, H., Lee, B. and Obrietan, K.: Mammalian Target of Rapamycin Signaling Modulates Photic Entrainment of the Suprachiasmatic Circadian Clock., J Neurosci 30, 6302, 2010 |
| 大鼠 | 大鼠, 食魚動物, 7-21 個月 | Hippocampal neurons | Papain: 0.2% | Hibernate A | Chen, N., Newcomb, J., Garbuzova-Davis, S., Davis Sanberg, C., Sanberg, P. and Willing, A.: Human Umbilical Cord Blood Cells Have Trophic Effects on Young and Aging Hippocampal Neurons in Vitro., Aging Dis 1, 173, 2010 |
| 大鼠 | 大鼠, SD, 7 天 | Cerebellar granule neurons | PDS kit: per instructions | PBS | Tanaka, S., Shaikh, I., Chiocca, E. and Saeki, Y.: The Gs-Linked Receptor GPR3 Inhibits the Proliferation of Cerebellar Granule Cells During Postnatal Development., PLoS ONE 4, e5922, 2009 |
| 大鼠 | 大鼠, neonatal | Astrocytes | Papain: 20 u/ml | EBSS | Shigetomi, E. and Khakh, B.: Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes., J Vis Exp 26, 1142, 2009 |
| 大鼠 | 大鼠,初生的 | Hippocampal neurons | Papain: 10 u/ml | EBSS | Richler Esther, Chaumont Severine, Shigetomi Eiji, Sagasti Alvaro, Khakh Baljit S: Tracking transmitter-gated P2X cation channel activation in vitro and in vivo,Nat Methods 5, 87-93, 2008 |
| 大鼠 | 大鼠, SD | Hippocampal | Trypsin: 0.1% | Neurobasal | Akanda, N., Tofighi, R., Brask, J., Tamm, C., Elinder, F. and Ceccali, S.:Voltage-Dependent Anion Channels (VDAC) in the Plasma Membrane Play a Critical Role in Apoptosis in Differentiated Hippocampal Neurons but not in Neural Stem Cells., Cell Cycle 7, 3225, 2008 |
| 大鼠 | 大鼠, neonatal | Dura mater | Collagenase Type 1: 0.075% | DMEM | Peptan, I., Hong, L. and Evans, C.: Multiple Differentiation Potentials of Neonatal Dura Mater-Derived Cells., Neurosurgery 60, 346, 2007 |
| 大鼠 | 大鼠, E18 | Hippocampal neurons | Papain: 0.2% | Hibernate | Jekabsons MB, Nicholls DG: Bioenergetic analysis of cerebellar granule neurons undergoing apoptosis by potassium/serum deprivation, Cell Death Differ 13, 1595-610, 2006 |
| 大鼠 | 大鼠, 威斯塔, 1-3 day | Hippocampal neurons | Trypsin: 0.05% | DMEM | Velasco Myrian, Garci璦 Esperanza, Onetti Carlos G: Glucose deprivation activates diversity of potassium channels in cultured rat hippocampal neurons,Cell Mol Neurobiol 26, 307-19, 2006 |
| 大鼠 | 大鼠, E19 | Hippocampal neuron | Papain: 10 u/ml | MEM | Khakh Baljit S, Fisher James A, Nashmi Raad, Bowser David N, Lester Henry A:An angstrom scale interaction between plasma membrane ATP-gated P2X2 and alpha4beta2 nicotinic channels measured with fluorescence resonance energy transfer and total internal reflection fluorescence microscopy, J Neurosci 25, 6911-20, 2005 |
| 大鼠 | 大鼠, 威斯塔, 14天 | Visual cortical | PDS kit: per instructions | EBSS | Mizoguchi Y, Kanematsu T, Hirata M, Nabekura J: A rapid increase in the total number of cell surface functional GABAA receptors induced by brain-derived neurotrophic factor in rat visual cortex, J Biol Chem 278, 44097, 2003 |
| 大鼠 | 大鼠, 威斯塔, 雄性 | Cerebral endothelial | Collagenase Type 3: 0.2% | MEM | Floris S, Van den Born J, van der Pol SM, Dijkstra CD, De Vries HE: Heparan sulfate proteoglycans modulate monocyte migration across cerebral endothelium, J Neuropathol Exp Neurol 62, 780, 2003 |
| 大鼠 | 大鼠, SD, E18 | Cortical cells | Papain: 20 u/ml | EBSS | Behar, T, Smith, S, Kennedy, R, Mckenzie, J, Maric, I and Barker, J: GABA(B) receptors mediate motility signals for migrating embryonic cortical cells, Cereb Cortex 11, 744-53, 2001 |
| Deoxyribonuclease I: .005% | |||||
| 大鼠 | 大鼠, 威斯塔, 250-300 g | Cerebral artery smooth muscle cells | Papain: 1.5 mg/ml | Physiological Salt Solution | Brzezinska AK, Gebremedhin D, Chilian WM, Kalyanaraman B, Elliott SJ.:Peroxynitrite reversibly inhibits Ca(2+)-activated K(+) channels in rat cerebral artery smooth muscle cells, Am J Physiol Heart Circ Physiol 278, H1883, 2000 |
| Collagenase Type 4: 1.5 mg/ml | |||||
| 大鼠 | 大鼠, SD, E16 | Cortical neurons | PDS kit: with modifications | Neurobasal Medium | Varney MA, Cosford ND, Jachec C, Rao SP, Sacaan A, Lin FF, Bleicher L, Santori EM, Flor PJ, Allgeier H, Gasparini F, Kuhn R, Hess SD, Velicelebi G, Johnson EC:SIB-1757 and SIB-1893: selective, noncompetitive antagonists of metabotropic glutamate receptor type 5, J Pharmacol Exp Ther 290, 170, 1999 |
| 大鼠 | 大鼠, 威斯塔, 初生的 | Hippocampal cells | Papain: 1 mg/ml | DMEM | Boehm S, Betz H: Somatostatin inhibits excitatory transmission at rat hippocampal synapses via presynaptic receptors, J Neurosci 17, 4066, 1997 |
| 大鼠 | 大鼠, SD, 食魚動物 | Hippocampal neurons | Papain: 0.2% | HibernateA/B27 | Brewer, G.J.: Isolation and culture of 成年的 rat hippocampal neurons of any age, J Neurosci Methods 71, 143, 1997 |
| 大鼠 | 大鼠, E18 | Hippocampal and cortical neurons | PDS kit: per instructions | Neurobasal media | Naeve GS, Ramakrishnan M, Kramer R, Hevroni D, Citri Y, Theill LE: Neuritin: a gene induced by neural activity and neurotrophins that promotes neuritogenesis,Proc Natl Acad Sci U S A 94, 2648-53, 1997 |
| 大鼠 | 大鼠, SD, 初生 | Glial | Trypsin: 0.0625% | MEM, sterile | Pixley, S.K.: The Olfactory Nerve Contains Two Populations of Glia, In Vitro 5, 269, 1992 |
| 大鼠 | 大鼠, 初生 | Astrocytes | Trypsin: 0.25% | DMEM, HBSS | Holzwarth, J., Glaum, S., and Miller, R.: Activation of Endothelin Receptors by Sarafotoxin Regulates Ca2+ Homeostasis in Cerebellar Astrocytes, Glia 5, 239, 1992 |
| 大鼠 | 大鼠, 60-72 小時大的 | Fibroblasts | Trypsin: 0.2% | HEPES buffered DMEM | Acheson, A., Barker, P., Alderson, R., Miller, F., and Murphy, R.: Detection of Brain-Derived Neurotrophic Factor-like Activity in Fibroblasts and Schwann Cells: Inhibition by Antibodies to NGF, Neuron 7, 265, 1991 |
| 大鼠 | 大鼠, SD, 成年的 | Endothelial | Collagenase Type 2: 0.5% | Medium 199 | Doron, D., Jacobowitz, D., Heldman, E., Feurerstein, G., Pollard, H., and Hallenbeck, J.: Extracellular Matrix Permits the Expression of Von Willebrand's Factor, Uptake of Di-I-Acetylated Low Density Lipoprotein and Secretion of Prostacyclin in Cultures of Endothelial Cells from Rat Brain Microvessels, In Vitro Cell Dev Biol 27A, 689, 1991 |
| 大鼠 | 大鼠, SD, 5天 | Hippocampal neurons | Trypsin: 0.2% | HBSS | Mattson, M., and Kater, S.: Development and Selective Neurodegeneration in Cell Cultures From Different Hippocampal Regions, Brain Res 490, 110, 1989 |
| Deoxyribonuclease I: 10 ug/ml | |||||
| 大鼠 | 大鼠, SD, 雌性 | Fetal rat brain | Collagenase Type 4: 0.1% | DMEM | Matsuda, M.: Serum Proteins Enhance Aggregate Formation of Dissociated Fetal Rat Brain Cells in an Aggregating Culture, In Vitro Cell Dev Biol 24 (10), 1031, 1988 |
| 大鼠 | 大鼠, 白化病, 成年的 和初生的 | Cerebral cortices | Trypsin: 0.25% | PBS | Giulian, D. and Baker, T.J.: Characterization of Ameboid Microglia Isolated From Developing Mammalian Brain, J Neurosci 6, 2163, 1986 |
| 大鼠 | 大鼠, 威斯塔, 初生 | Germinal matrix | Trypsin: 0.25% | HBSS | Goldman, J.E., Geier, S.S., and Hirano, M.: Differentiation of Astrocytes and Oligodendrocytes From Germinal Matrix Cells in Primary Culture, J Neurosci 6, 52, 1986 |
| 大鼠 | 大鼠,胎兒, 18-20天 | Hippocampal neurons | Trypsin: 0.25% | HBSS, CMF | Bartlett, W. and Banker, G.: An Electron Microscopic Study of the Development of Axons and Dendrites by Hippocampal Neurons in Culture. I. Cells Which Develop Without Intercellular Contacts, J Neurosci 4, 1944, 1984 |
| 大鼠 | 大鼠, 胎兒, 18 天 | Hippocampi | Trypsin: 0.1% | HBSS | Rothman, S.: Synaptic Release of Excitatory Amino Acid Neurotransmitter Mediates Anoxic Neuronal Death, J Neurosci 4 (7), 1884, 1984 |
| 大鼠 | 大鼠, SD, 懷孕19-20天的 | Neural | Trypsin: 0.25% | DMEM | Ahmed, Z., Walker, P., and Fellows, R.: Properties of Neurons from Dissociated Fetal Rat Brain in Serum-Free Culture, J Neurosci 3 (12), 2448, 1983 |
| 大鼠 | 大鼠, SD, 胎兒 | Cerebral cortex | Deoxyribonuclease I: 0.001% | HEPES | Peterfreund, R. and Vale, W.: High Molecular Weight Somatostatin Secretion by Cultured Rat Brain Cell, Brain Res 239, 463, 1982 |
| Hypothalmus | |||||
| 大鼠 | Wistar京都大鼠, 雄性, 100 - 200 克 | Endothelial | Collagenase Type 2: 0.05% | HBSS | Diglio, C.A., Grammas, P., Filiberto Giacomelli, M.S., and Wiener, J.: Primary Culture of Rat Cerebral Microvascular Endothelial Cells, Lab Invest 46, 554, 1982 |
| Cerebral | |||||
| 大鼠 | 大鼠, 胎兒 | Cortical | Trypsin: 0.25% | Puck's D1 (see reference) | Swaiman, K., Neale, E., Fitzgerald, S., and Nelson, P.: A Method for Large-scale Production of Mouse Brain Cortical Cultures, Brain Res 255, 361, 1982 |
| 大鼠 | 大鼠, 威斯塔,懷孕的 | Glial | Trypsin: 0.05% | Eagle's MEM/DMEM | Abney, E., Bartlett, P., and Raff, M.: Astrocytes, Ependymal Cells, and Oligodendrocytes Develop on Schedule in Dissociated Cell Cultures of Embryonic Rat Brain, Dev Biol 83, 301, 1981 |
| 大鼠 | 大鼠, SD, 一個月大的 | Capillary endothelium | Neutral Protease: 0.1% | Medium 199 | Bowman, P., Betz, A., Ar, D., Wolinsky, J., Penney, J., Shivers, R., and Goldstein, G.: Primary Culture of Capillary Endothelium From Rat Brain, In Vitro 17 (4), 353, 1981 |
| Pericytes | |||||
| 大鼠 | 大鼠, 威斯塔, 雄性, 300-500 克 | Microvessels | Collagenase Type 2: 0.75% | Ringers-HEPES buffer | Williams, S., Gillis, J., Matthews, M., Wagner, R., and Bitensky, M.: Isolation and Characterization of Brain Endothelial Cells: Morphology and Enzyme Activity, J Neurochem 35 (2), 374, 1980 |
| Endothelial | |||||
| 大鼠 | 大鼠, 威斯塔, 成年的, 170克 | Endothelial | Trypsin: 0.5% | BSS | Phillips, P., Kumar, P., Kumar, S., and Waghe, M.: Isolation And Characterization of Endothelial Cells From Rat And Cow Brain White Matter, J Anat 129, 261, 1979 |
| 大鼠 | 大鼠, 豪斯曼, 18 天 | Hippocampal neurons | Trypsin: 0.1% | HBSS | Banker, G., and Cowan, M.: Rat Hippocampal Neurons in Dispersed Cell Culture,Brain Res 126, 397, 1977 |
| 大鼠 | 大鼠, SD, 2 天 | Pineal | Trypsin: 0.25% | DMEM, MEM | Rowe, V., Neale, E., Avins, L., Guroff, G., and Schrier, B.: Pineal gland cells in culture. Morphology, Biochemistry, Differentiation, and co-culture with sympathetic neurons, Exp Cell Res 104, 345, 1977 |
| 貝類動物 | 旋節(jié)螺trivolvis | Neurons | Trypsin: 0.2% | Saline, Sterile | Hadley, R.D., Bodnar, D.A., and Kater, S.B.: Formation of Electrical Synapses Between Isolated, Cultured Helisoma Neurons Requires Mutual Neurite Elongation,J Neurosci 5, 3145, 1985 |
| Buccal ganglia |
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