国产三级毛片麻豆-国产精品无码一区二区三区免费-国产麻豆午夜三级精品-亚洲国产日韩?在线播放-精品国精品国产自在久国产应用-国产精品久久久中文字幕-麻豆AV无码久久精品蜜桃久久

新聞中心

News Center

當前位置:首頁新聞中心神經(jīng)組織消化手冊

神經(jīng)組織消化手冊

更新時間:2016-02-16點擊次數(shù):1971

世界*實驗材料供應商 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,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,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,您的滿意將是我們zui大的收獲

 Worthington Biochemical 中國代理, Worthington Biochemical 上海代理, Worthington Biochemical 北京代理,Worthington Biochemical 廣東代理, Worthington Biochemical 江蘇代理Worthington Biochemical 湖北代理,Worthington Biochemical 天津,Worthington Biochemical 黑龍江代理,Worthington Biochemical 內(nèi)蒙古代理,Worthington Biochemical 吉林代理,Worthington Biochemical 福建代理, Worthington Biochemical 江蘇代理, Worthington Biochemical 浙江代理,Worthington Biochemical 四川代理,

 

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)具體實驗目的修正消化方案以獲得更好的實驗結(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,描述了標準的實驗步驟, 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

心臟

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。

 

神經(jīng)組織消化手冊

 

物種詳細來源細胞名稱消化用酶培養(yǎng)基參考文獻
鳥類小雞, 白來杭雞, 胚胎Cilary ganglion neuronsCollagenase Type 1: 0.1%PBSTemburni, M., Rosenberg, M., Pathak, N., McConnell, R. and Jacob, M.:Neuronal Nicotinic Synapse Assembly Requires the Adenomatous Polyposis Coli Tumor Suppressor Protein., J Neurosci 24, 6776, 2004
牛 HeartTrypsin: 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
(包括 大鼠)Adrenal chromaffin 
 Paraneurons
Microvascular endothelialCollagenase/Dispase: 0.1%MEMBowman, P., Betz, A., and G大stein, G.: Primary Culture of Microvascular Endothelial Cells From Bovine Retina, In Vitro 18 (7), 626, 1982
牛犢OligodendrogliaTrypsin: 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
(包括 小羊羔)Neural
雞, 白來杭雞, 胚胎, 17-21 天NeuronsPapain: 40 u/mlHEPESRaman, I., and Trussell, L.: The Kinetics of the Response to Glutamate and Kainte in Neurons of the Avian and Cochlear Nucleus, Neuron 9, 173, 1992
小雞, 胚胎, 10-14 天Flat, retinaTrypsin: 0.1%Tyrode's solution, CMFMoyer, M., Bullrich, F., and Sheffield, J.: Emergence of Flat Cells From Glia in Stationary Cultures of 胚胎Chick Neural Retina, In Vitro Cell Dev Biol26, 1073, 1990
小雞, 白來杭雞, 胚胎Cerebral neuronsTrypsin: Trypsin: 0.125-0.25%DMEMCoates, P., and Nathan, R.: Feasibility of Electrical Recordings From Unconnected Vertebrate CNS Neurons Cultured in a Three-Dimensional Extracellular Matrix, J Neurosci Methods 20, 203, 1987
小雞, 白來杭雞, 胚胎Spinal cordTrypsin: 0.05%Phosphate bufferSchnaar, R., and Schaffner, A.: Separation of cell types from 胚胎chicken and rat spinal cord: characterization of motoneuron-enriched fractions, J Neurosci 1, 204, 1981
(包括SD大鼠, 胚胎)(see reference)
小雞Dorsal root ganglion neuronsTrypsin: 0.1%Puck's saline, CMFChoi, D., and Fiscbach, G.: GABA Conductance of Chick Spinal Cord and Dorsal Root Ganglion Neurons in Cell Culture, J Neurophysiol 45, 605, 1981
Spinal cord
小雞, 白來杭雞 胚胎, 8 天Neurons, gangliaTrypsin: 0.25%HBSS, CMFBottenstein, J., Skaper, S., Varon, S., and Sato, G.: Selective Survival of Neurons from Chick 胚胎 Sensory Ganglionic Dissociates Utilizing Serum-Free Supplemented Medium, Exp Cell Res 125, 183, 1980
雞, 白來杭雞, 種蛋Ciliary ganglion neuronsTrypsin: 0.25%Eagle's MEMTuttle, J., Suszkiw, J., and Ard, M.: Long-Term Survival and Development of Dissociated Parasympathetic Neurons in Culture, Brain Res 183, 161, 1980
雞, 胚胎, 9-10 天大Dorsal root ganglia neuronsCollagenase: 0.01%Eagle's MEMMudge, A., Leeman, S., and Fischbach, G.: Enkephalin Inhibits Release of Substance P From Sensory Neurons in Culture and Decreases Action Potential Duration, Proc Natl Acad Sci U S A 76 (1), 526, 1979
小雞胚胎Ganglion chains, sympathetic gangliaTrypsin: 0.25%Krebs PhosphosalineMcCarthy, K., and Partlow, L.: Preparation of Pure Neuronal and Non-Neuronal Cultures From 胚胎Chick Sympathetic Ganglia: A New Method Based on Both Differential Cell Adhesiveness and the Formation of Homotypic Neuronal Aggregates, Brain Res 114, 391, 1976
小雞, 白來杭雞, 胚胎, 7 天Neural retinaCollagenase: 0.25%HBSSWiseman, L., and Hammond, W.: The Reacquisition of Cell Adhesiveness Following Tissue Disaggregation by Eleven Different Agents, J Exp Zool 197, 429, 1976
Elastase: 0.2%
Hyaluronidase: 1.0%
Papain: 1.0%
Protease: 0.1%
Trypsin: 0.05%
魚, 斑馬魚Motor neuronsCollagenase Type 2: 0.1%See referenceSakowski, S., Lunn, J., Busta, A., Palmer, M., Dowling, J. and Feldman, E.: A Novel Approach to Study Motor Neurons from Zebrafish 胚胎s and Larvae in Culture., J Neurosci Methods 205, 277-82, 2012
 Dania rerio斑馬魚,  胚胎Rohon-Beard neuronsTrypsin: 0.2%L-15/HepesWon, Y., Ono, F. and Ikeda, S.: Identification and Modulation of Voltage-Gated Ca2+ Currents in Zebrafish Rohon-Beard Neurons., J Neurophysiol 105, 442-53, 2011
Dania rerio斑馬魚, 胚胎NeuronsPDS kit: with modificationsEBSSCerda, G., Hargrave, M. and Lewis, K.: RNA Profiling of FAC-sorted Neurons from the Developing Zebrafish Spinal Cord., Dev Dyn 238, 150-61, 2009
黑色的魅影kni克e魚, 成年的(黑魔鬼魚)Neurons, spinal cordTrypsin: 0.4%PBS, CMFAnderson, M.J.: Differences in Growth of Neurons from Normal and Regenerated eost Spinal Cord in vitro, In Vitro Cell Dev Biol 29A, 145, 1993
青蛙非洲爪蟾蜍, 胚胎NeuronCollagenase Type 1: 0.1%Steinberg's solutionTakahashi, T., Nakajima, Y., Hirosawa, K., Nakajima, S., and Onodera, K.: Structure and Physiology of Developing Neuromuscular Synapses in Culture, J Neurosci 7, 473, 1987
豚鼠豚鼠, 初生Neuron, entericTrypsin: 0.125%Medium 199Jessen, K, McConnell, J., Purves, R., Burnstock, G., and Chamley-Campbell, J.: Tissue Culture of Mammalian Enteric Neurons, Brain Res 152, 573, 1978
馬, 雄性Vomeronasal organ neuronsCollagenase Type 1: 0.02%PBSLiman ER: Regulation by voltage and adenine nucleotides of a Ca2+-activated cation channel from hamster vomeronasal sensory neurons, J Physiol 548, 777, 2003
Trypsin: 0.02%
人類人類Neural, variousPapain: 12 u/mlsee referencePanchision David M, Chen Hui-Ling, Pistollato Francesca, Papini Daniela, Ni Hsiao-Tzu, Hawley Teresa S: Optimized flow cytometric analysis of central nervous system tissue reveals novel functional relationships among cells expressing CD133, CD15, and CD24, Stem Cells 25, 1560-70, 2007
Trypsin: see reference
Collagenase/Dispase: see reference
人類人類, 9 個月NeuronsCollagenase Type 4: 1.33%DMEM/F12Dietrich J, Lacagnina M, Gass D, Richfield E, Mayer-Proschel M, Noble M, Torres C, Proschel C.: EIF2B5 mutations compromise GFAP+ astrocyte generation in vanishing white matter leukodystrophy, Nat Med 11, 277, 2005
Papain: 0.07 u/ml
Neutral Protease: 1 mg/ml
人類人類, 26 周Neural progenitor cellsPapain: 2.5 u/mlDMEM/F-12Fuja TJ, Schwartz PH, Darcy D, Bryant PJ.: Asymmetric localization of LGN but not AGS3, two homologs of Drosophila pins, in dividing human neural progenitor cells, J Neurosci Res 75, 782, 2004
Deoxyribonuclease I: 250 u/ml
Neutral Protease: 1 u/ml
人類人類, 成年人Ventricular epithelialPapain: 11.4 u/mlDMEM/F12Roy NS, Benraiss A, Wang S, Fraser RA, Goodman R, Couldwell WT, Nedergaard M, Kawaguchi A, Okano H, G大man SA: Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone, J Neurosci Res 59, 321, 2000
Deoxyribonuclease I: 10 u/ml
人類人類, 5-65 歲Retinal pigment epithelial (RPE)Trypsin: 0.25%HBSSVon Recum, H., Okano, T., Kim, S, and Bernstein, P.: Maintenance of Retinoid Metabolism in Human Retinal Pigment Epithelium Cell Culture, Exp Eye Res 69, 97, 1999
人類人類Dorsal root ganglion neuronsCollagenase Type 1: 0.2%DMEM/F-12Dib-Hajj SD, Tyrrell L, Cummins TR, Black JA, Wood PM, Waxman SG.:Two tetrodotoxin-resistant sodium channels in human dorsal root ganglion neurons, F.E.B.S. Lett. 462, 117, 1999
Neutral Protease: 0.5%
昆蟲果蠅, 幼蟲NeuroblastsCollagenase Type 1: 0.1%Rinaldini solutionBerger, C., Harzer, H., Burkard, T., Steinmann, J., van der Horst, S., Laurenson, A., Novatchkova, M., Reichert, H. and Knoblich, J.: FACS Purification and Transcriptome Analysis of Drosophila Neural Stem Cells Reveals a Role for Klumpfuss in Self-Renewal., Cell Rep 2, 407, 2012
Papain: 0.1%
昆蟲果蠅DendritesCollagenase: 0.05%HBSSSanchez-Soriano, N., Bottenberg, W., Fiala, A., Haessler, U., Kerassoviti, A., Knust, E., Lohr, R. and Prokop, A.: Are Dendrites in Drosophila Homologous to Vertebrate Dendrites?, Dev Biol 288, 126, 2005
Neutral Protease: 0.2%
昆蟲雙斑蟋Giant interneuronsCollagenase: 0.05%Leibovitz's L15Kloppenburg, P. and Horner, M.: Voltage-Activated Currents in Identified Giant Interneurons Isolated from Adult Crickets Gryllus Bimaculatus, J Exp Biol201 (Pt 17), 2529, 1998
Neutral Protease: 0.2%
小鼠小鼠DRG neuronsCollagenase Type 2: 0.1%DMEM/F-12Lee, B., Cho, H., Jung, J., Yang, Y., Yang, D. and Oh, U.: Anoctamin 1 Contributes to Inflammatory and Nerve-Injury Induced Hypersensitivity., Mol Pain 10, 5, 2014
Trypsin: 0.25%
小鼠小鼠, 新生的Schwann cellsCollagenase: 0.05-0.1%DMEMStettner, M., Lohmann, B., Wolffram, K., Weinberger, J., Dehmel, T., Hartung, H., Mausberg, A. and Kieseier, B.: Interleukin-17 Impedes Schwann Cell-Mediated Myelination., J Neuroinflammation 11, 63, 2014
Trypsin: 0.125-0.25%
小鼠小鼠OligodendrocytesPapain: 0.1%HBSSHarris, M., Hulseberg, P., Ling, C., Karman, J., Clarkson, B., Harding, J., Zhang, M., Sandor, A., Christensen, K., Nagy, A., Sandor, Matyas,S. and Fabry, Z.: Immune Privilege of the CNS is not the Consequence of Limited Antigen Sampling., Sci Rep 4, 4422, 2014
Deoxyribonuclease I: 0.0002%
小鼠小鼠hippocampal neuronsPDS kit: per instructionsDMEMWilliams, E., Zhong, X., Mohamed, A., Li, R., Liu, Y., Dong, Q., Ananiev, G., Mok, J., Lin, B., Lu, J., Chiao, C., Cherney, R., Li, H., Zhang, S. and Chang, Q: Mutant Astrocytes Differentiated from Rett Syndrome Patients-Specific iPSCs Have Adverse Effects on Wild-Type Neurons., Hum Mol Genet23, 2968-80, 2014
小鼠小鼠NeurosperesPapain: 0.1%NeuroCult NSCStuart, M., Corrigan, F. and Baune, B.: Knockout of CXCR5 Increases the Population of Immature Neural Cells and Decreases Proliferation in the Hippocampal Dentate Gyrus., J Neuroinflammation 11, 31, 2014
Deoxyribonuclease I: 0.1%
小鼠小鼠, 4-6 周SchwannCollagenase Type 1: 0.01%DMEMMasaki, T., Qu, J., Cholewa-Waclaw, J., Burr, K., Raaum, R. and Rambukkana, A.: Reprogramming Adult Schwann Cells to Stem Cell-Like Cells by Leprosy Bacilli Promotes Dissemination of Infection., Cell 152, 51, 2013
Trypsin: 0.125%
小鼠小鼠Oligodendrocyte precursor cellsPapain: 0.09%DMEMMedina-Rodriguez, E., Arenzana, F., Bribian, A. and de Castro, F.:Protocol to Isolate a Large Amount of Functional Oligodendrocyte Precursor Cells from the Cerebral Cortex of Adult Mice and Humans., PLoS ONE 8, e81620, 2013
小鼠小鼠, 6-8 周Neural precursorsTrypsin: 0.13%see referenceBabona-Pilipos, R., Popovic, M. and Morshead, C.: A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field., J Vis Exp 68, e4193, 2012
Hyaluronidase: 0.08%
小鼠小鼠, 成年的Retinal neuronsPapain: 10-20 ulHBSSGoetz, J. and Trimarchi, J.: Single-Cell Profiling of Developing and Mature Retinal Neurons., J Vis Exp 62, e3824, 2012
小鼠小鼠NeuronsPapain: 100 ulDMEMLi, Y., Roy, B., Wang, W., Zhang, L., Sampson, S. and Lin, D.: Imaging pHluorin-Tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons., J Vis Exp 69, e4450, 2012
小鼠小鼠, P4-5NeuroshereTrypsin: 0.25%Pro-NZiegler, A., Schneider, J., Qin, M., Tyler, W., Pintar, J., Fraidenraich, D., Wood, T and Levison, S.: IGF-II Promotes Stemness of Neural Restricted Precursors., Stem Cells 30, 1265, 2012
Papain: 100 u
Deoxyribonuclease I: 0.025%
小鼠小鼠, 6-8 周Neural stem cellsPapain: 0.01%DMEM/F12Bracko, O., Singer, T., Aigner, S., Knobloch, M., Winner, B., Ray, J., Clemenson, G., Suh, H., Couillard-Despres, S., Aigner, L., Gage, F. and Jessberger, S.: Gene Expression Profiling of Neural Stem Cells and their Neuronal Progeny Reveals IGF2 as a Regulator of Adult Hippocampal Neurogenesis., J Neurosci 32, 3376-87, 2012
Neutral Protease: 0.1%
Deoxyribonuclease I: 0.01%
小鼠小鼠, 胚胎, 14 天Spinal cord neuronsPapain: 0.05%PBS/DMEMPollari, E., Savchenko, E., Jaronen, M., Kanninen, K., Malm, T., Wojciechowski, S., Ahtoniemi, T., G大steins, G., Giniatullina, R., Giniatullin, R., Koistinaho, J. and Magga, J.: Granulocyte Colony Stimulating Factor Attenuates Inflammation in a Mouse Model of Amyotrophic Lateral Sclerosis., J Neuroinflammation 8, 74, 2011
Deoxyribonuclease I: 0.004%
小鼠小鼠, 1-2 天Oligodendrocytes, dorsal root gangliaPapain: 0.15%DMEMO'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, 胚胎MotorneuronsTrypsin: 0.025%HBSSConrad, R., Jablonka, S., Sczepan, T., Sendtner, M., Wiese, S. and Klausmeyer, A.: Lectin-Based Isolation and Culture of Mouse 胚胎Motoneurons., J Vis Exp 55, e3200, 2011
小鼠小鼠, 4-8 周Olfactory ensheathingCollagenase: 0.15%DMEMRadtke, C., Sasaki, M., Lankford, K., Gallo, V. and Kocsis, J.: CNPase Expression in Olfactory Ensheathing Cells., J Biomed Biotechnol 2011, 608496, 2011
Papain: 12 u/ml
小鼠小鼠Dentate Gyrus Neural PrecursorPapain: 2.5 u/mlNeurobasal ABabu, H., Claasen, J., Kannan, S., Runker, A., Palmer, T. and Kempermann, G.: A Protocol for Isolation and Enriched Monolayer C*tion of Neural Precursor Cells from Mouse Dentate Gyrus.,Front Neurosci Vol. 5,, , 89, 2011
Neutral Protease: 1.0 u/ml
Deoxyribonuclease I: 250 u/ml
小鼠小鼠Hippocampal and retinal neuronsPapain: 1%HBSSBrown, J., Gianino, S. and Gutmann, D.: Defective cAMP Generation Underlies the Sensitivity of CNS Neurons to Neurofibromatosis-1 Heterozygosity, J Neurosci 30, 5579, 2010
Deoxyribonuclease I: 5 u/ml
小鼠小鼠CNS leukocytesCollagenase Type 4: 300 u/mlHBSSSayed, B., Christy, A., Walker, M. and Brown, M.: Meningeal Mast Cells Affect Early T Cell Central Nervous System Infiltration and Blood-Brain Barrier Integrity Through TNF: a Role for Neutrophil Recruitment?, J Immunol 184, 6891-900, 2010
小鼠小鼠, 成年的Spinal microgangliaPapain: 0.2%Hibernate AYip, P., Kaan, T., Fenesan, D. and Malcangio, M.: Rapid Isolation and Culture of Primary Microglia from Adult Mouse Spinal Cord., J Neurosci Methods 183, 223-37, 2009
小鼠小鼠, 胚胎Dopaminergic neuronsTrypsin: 0.1%DMEMRadad Khaled, Gille Gabriele, Rausch Wolf-Dieter: Dopaminergic neurons are preferentially sensitive to long-term rotenone toxicity in primary cell culture, Toxicol In Vitro 22, 68-74, 2008
Deoxyribonuclease I: 0.02%
小鼠小鼠, 新生的DRG neuronsCollagenase: 0.2%Ham's F12Pedrola, L., Espert, A., Valdes-Sanchez, T., Sanchez-Piris, M., Sirkowski, E., Scherer, S., Farinas, I., and Palau, F.: Cell Expression of GDAP1 in the Nervous System and Pathogenesis of Charcot-Marie-Tooth Type 4A Disease.,J Cell Mol Med 12, 679, 2008
Trypsin: 0.05%
小鼠小鼠, 6 個月Neurons, neurospheresPapain: 0.2%HibernateBrewer Gregory J, Torricelli John R: Isolation and culture of adult neurons and neurospheres, Nat Protoc 2, 1490-8, 2007
小鼠小鼠Neural, variousPapain: 12 u/mlsee referencePanchision David M, Chen Hui-Ling, Pistollato Francesca, Papini Daniela, Ni Hsiao-Tzu, Hawley Teresa S: Optimized flow cytometric analysis of central nervous system tissue reveals novel functional relationships among cells expressing CD133, CD15, and CD24, Stem Cells 25, 1560-70, 2007
Trypsin: see reference
Collagenase/Dispase: see reference
小鼠小鼠, 6-8 周Sensory neurons, DRGPapain: 20 u/mlHBSSMalin, S., Davis, B. and Molliver, D.: Production of Dissociated Sensory Neuron Cultures and Considerations for their use in Studying Neuronal Function and Plasticity, Nat Protoc 2, 152, 2007
Collagenase Type 2: 0.4%
Neutral Protease: 0.46%
小鼠小鼠, C57BL/6, 1 歲NeuronsPapain: 0.2%DMEMEide, L, and McMurray, C: Culture of Adult Mouse Neurons, Biotechniques38(1), 99-104, 2005
小鼠小鼠, 成年的Brain and spinal cord cellsTrypsin: 0.25%PBSGonzalez John M, Bergmann Cornelia C, Fuss Babette, Hinton David R, Kangas Cindy, Macklin Wendy B, Stohlman Stephen A: Expression of a dominant negative IFN-gammareceptor on mouse oligodendrocytes, Glia 51, 22-34, 2005
小鼠小鼠, 新生的NeuronsPDS kit: per instructionsEBSSGill JC, Moenter SM, Tsai PS: Developmental regulation of gonadotropin-releasing hormone neurons by fibroblast growth factor signaling,Endocrinology 145, 3830, 2004
小鼠小鼠, C57BLCerebellar granule cell precursorsPapain: 16.5 u/mlDulbecco's PBSOkano-Uchida T, Himi T, Komiya Y, and Ishizaki Y: Cerebellar granule cell precursors can differentiate into astroglial cells, Proc Natl Acad Sci U S A 101, 1211, 2004
Deoxyribonuclease I: 0.008%
小鼠小鼠, 1-3 天Neurons, gangliaPapain: 20 u/mlHBSSSavchenko V, Sung U, Blakely RD.: Cell surface trafficking of the antidepressant-sensitive norepinephrine transporter revealed with an ectodomain antibody, Mol Cell Neurosci 24, 1131, 2003
Deoxyribonuclease I: 100 u/ml
Collagenase: 0.3%
Trypsin: 0.05%
小鼠小鼠,雄性Trigeminal senesory neuronsPapain: 20 u/mlHEPES buffered salineRoberts, L., MacDonald, C.and Mark, C.: Anandamide is a Partial Agonist at Native Vanilloid Receptors in Acuy Isolated Mouse Trigeminal Sensory Neurons., Br J Pharmacol 137, 421, 2002
小鼠小鼠, 出生后一天的NeuronsTrypsin: 0.25%NGF-containing mediumDeshmukh, M., Kuida, K., and Johnson Jr., E.: Caspase Inhibition Extends the Commitment to Neuronal Death Beyond Cytochrome c Release to the Point of Mitochondrial Depolarization, J Cell Biol 150 (1), 131, 2000
小鼠小鼠 胚胎Neurons, DRG and SCGTrypsin: 0.25%L-15 mediumLee, K., Davies, A., and Jaenisch, R.: P75-Eficient 胚胎Dorsal Root Sensory and Neonatal Sympathetic Neurons Display a Decreased Sensitivity to NGF, Development 120, 1027, 1994
小鼠小鼠, 65 天Neurons, neuronal precursorsTrypsin: 0.1%DMEMRichards L., Kilpatrick T., and Bartlett P.: De Novo Generation of Neuronal Cells from the Adult Mouse Brain, Proc Natl Acad Sci U S A 89, 8591, 1992
Deoxyribonuclease I: 0.001%
小鼠小鼠, CD-1, neonateNeurons, dorsal root ganglionTrypsin: 0.25%HBSSQuinn, S. and De Boni, U.: Enhanced Neuronal Regeneration by Retinoic Acid of Murine Dorsal Root Ganglia and of Fetal Murine and human Spinal Cord in vitro, In Vitro Cell Dev Biol 27, 55, 1991
小鼠小鼠, 胎兒PrecursorTrypsin: 0.5%PBSKitani, H., Shiurba, R., Sakakura, T., Tomooka, Y.: Isolation and Characterization Of Mouse Neural Precursor Cells in Primary Culture, In Vitro Cell Dev Biol 27, 615, 1991
小鼠小鼠 (SWR or CF1), 1-3 月sPapillae, taste receptorPronase E: 0.15%Carbonate-Phosphate bufferSpielman, 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, 新生的PNS test neuronsTrypsin: 0.08%Eagle's Basal Medium (see reference)Varon, S., Skaper, S., Barbin, G., Selak, I., and Manthorpe, M.: Low Molecular Weight Agents Support Survival of Cultured Neurons From the Central Nervous System, J Neurosci 4 (3), 654, 1984
(包括 小雞)
小鼠小鼠 (白變種實驗室老鼠), 成年的Neurons, spinal cordCollagenase Type 3: 0.25%Hank's BSS, CMFEagleson, K. and Bennett, M.: Survival of Purified Motor Neurons In Vitro: Effects of Skeletal Muscle-Conditioned Medium, Neurosci Lett 38, 187, 1983
小鼠小鼠, 0-30 天NeuralTrypsin NF 1:250: 50 0.25%BSSShrier, B., Wilson, S., and Nirenberg, M.: Cultured Cell Systems and Methods for Neurobiology, Vol. 32,, , 765, 1974
綿羊羊羔OligodendrogliaTrypsin: 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
(包括 小牛)Neural
豬, 成年的, 60-100 公斤Superior cervical gangliaPapain: 2 u/mlHBSSSi ML, Lee TJ.: Presynaptic alpha7-nicotinic acetylcholine receptors mediate nicotine-induced nitric oxidergic neurogenic vasodilation in porcine basilar arteries, J Pharmacol Exp Ther 298, 122, 2001
Collagenase: 0.12%
Neutral Protease: 0.48%
鵪鶉鵪鶉Neural crestTrypsin: 0.05%MEM, HBSSSieber-Blum, M., and Cohen, A.: Clonal Analysis of Quail Neural Crest Cells: They are Pluripotent and Differentiate in Vitro in the Absence of Noncrest Cells, Dev Biol 80, 96, 1980
大鼠大鼠, 胚胎和 新生的Cortical neurons, DRGPDS kit: per instructionsNeurobasalRheaume, C., Cai, B., Wang, J., Fernandez-Salas, E., Aoki, K., Francis, J. and Broide, R.: A Highly Specific Monoclonal Antibody for Botulinum Neurotoxin Type A-Cleaved SNAP25., Toxins 7, 2354-70, 2015
Collagenase Type 1: 0.17%
大鼠大鼠, 胚胎Cerebral neuronsPapain: 2 u/mlDMEM-PBSOyanagi, K., Tashiro, T. and Negishi, T.: Cell-Type-Specific and Differentiation-Status-Dependent Variations in Cytotoxicity of Tributyltin in Cultured Rat Cerebral Neurons and Astrocytes., J Toxicol Sci 40, 459-68, 2015
Deoxyribonuclease I: 0.01%
大鼠SD大鼠, 6-9 周Dorsal Root GangliaCollagenase Type 3: 0.2%HBSSMurayama, C., Watanabe, S., Nakamura, M. and, Norimoto, H.: Inhibitory Activity of Yokukansankachimpihange against Nerve Growth Factor-Induced Neurite Growth in Cultured Rat Dorsal Root Ganglion Neurons.,Molecules 20, 14959-69, 2015
大鼠大鼠Dorsal root ganglion neuronsCollagenase Type 2: 0.15%HBSSYu, R., Seymour, V., Berecki, G., Jia, X., Akca, M., Adams, D., Kaas, Q. and Craik, D.: Less is More: Design of a Highly Stable Disulfide-Deleted Mutant of Analgesic Cyclic 偽-Conotoxin Vc1.1., Sci Rep 5, 13264, 2015
大鼠大鼠, 新生的Schwann cellsCollagenase: 0.05-0.1%DMEMStettner, M., Lohmann, B., Wolffram, K., Weinberger, J., Dehmel, T., Hartung, H., Mausberg, A. and Kieseier, B.: Interleukin-17 Impedes Schwann Cell-Mediated Myelination., J Neuroinflammation 11, 63, 2014
Trypsin: 0.125-0.25%
大鼠大鼠, 1-3 天Hippocampal NeuronsPapain: 25 u/mlL-15Thurner, P., Gsandtner, I., Kudlacek, O., Choquet, D., Nanoff, C., Freissmuth, M. and Zezula, J.: A Two-State Model for the Diffusion of the A2A Adenosine Receptor in Hippocampal Neurons: Agonist-Induced Switch to Slow Mobility is Modified by Synapse-Associated Protein 102 (SAP102)., J Biol Chem 289, 9263-74, 2014
大鼠SD大鼠, 胚胎Hypothalamic neuronalPDS kit: per instructionsNeurobasal ALoktev, A. and Jackson, P.: Neuropeptide Y Family Receptors Traffic via the Bardet-Biedl Syndrome Pathway to Signal in Neuronal Primary Cilia., Cell Rep 5, 1316-29, 2013
大鼠SD大鼠, 175-200 克Trigeminal gangliaPapain: 20 u/mlHBSSYan, J., Melemedjian, O., Price, T. and Dussor, G.: Sensitization of Dural Afferents Underlies Migraine-Related Behavior Following Meningeal Application of Interleukin-6 (IL-6)., Mol Pain 8, 6, 2012
Collagenase Type 2: 0.3%
大鼠大鼠, E19-21 and P1-3Sympathetic neuronsCollagenase Type 2: 0.1%DMEM/F-12Ghogha, A., Bruun, D. and Lein, P.: Inducing Dendritic Growth in Cultured Sympathetic Neurons., J Vis Exp 61, e3546, 2012
Neutral Protease: 0.5%
大鼠大鼠, 20-30 天Dorsal Root GanglionPapain: 20 u/mlL-15Bosmans, F., Puopolo, M., Martin-Eauclaire, M., Bean, B., Swartz, K.:Functional Properties and Toxin Pharmacology of a Dorsal Root Ganglion Sodium Channel Viewed through its Voltage Sensors., J Gen Physiol 138, 59, 2011
Collagenase Type 1: 0.3%
Neutral Protease: 0.4%
大鼠SD大鼠, 200-300 克Dorsal root ganglia, fibroblastCollagenase: 0.125%DMEMEast, E., de Oliveira, D., G大ing, J. and Phillips, J.: Alignment of Astrocytes Increases Neuronal Growth in Three-Dimensional Collagen Gels and is Maintained Following Plastic Compression to Form a Spinal Cord Repair Conduit., Tissue Eng Part A 16, 3173, 2010
大鼠大鼠,食魚貂, 7-21 月Hippocampal neuronsPapain: 0.2%Hibernate AChen, 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SD大鼠, E18NeuronalPapain: 0.2%Neurobasal EPeltier, D., Simms, A., Farmer, J and Miller, D.: Human Neuronal Cells Possess Functional Cytoplasmic and TLR-Mediated Innate Immune Pathways Influenced by Phosphatidylinositol-3-Kinase Signaling, J Immunol 184, 7010, 2010
大鼠SD大鼠, 7 天Cerebellar granule neuronsPDS kit: per instructionsPBSTanaka, 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SD大鼠, 胚胎天 18Hippocampal neuronsPapain: 20 u/mlNeurobasal/B27Liu, Y., Yohrling, G., Wang, Y., Hutchinson, T., Brenneman, D., Flores, C., Zhao, B.: Carisbamate, a Novel Neuromodulator, Inhibits Voltage-Gated Sodium Channels and Action Potential Firing of Rat Hippocampal Neurons.,Epilepsy Res 83, 66, 2009
大鼠大鼠, 胚胎Hypothalamic neurons and gliasPapain: 0.5 u/mlDMEMYokosuka Makoto, Ohtani-Kaneko Ritsuko, Yamashita Kayoko, Muraoka Daisuke, Kuroda Yoichiro, Watanabe Chiho: Estrogen and environmental estrogenic chemicals exert developmental effects on rat hypothalamic neurons and glias, Toxicol In Vitro 22, 1-9, 2008
大鼠 Wistar大鼠, 7 天Superior cervical ganglionCollagenase: 0.05%L-15Sakisaka, T., Yamamoto, Y., Mochida, S., Nakamura, M., Nishikawa, K., Ishizaki, H., Okamoto-Tanaka, M., Miyoshi, J., Fujiyoshi, Y., Manabe, T. and Takai, Y.: Dual Inhibition of SNARE Complex Formation by Tomosyn Ensures Controlled Neurotransmitter Release., J Cell Biol 183, 323, 2008
大鼠大鼠, 新生的Dopamine neuronsPapain: 20 u/mlPBSFrank, L., Caldera-Siu, A. and Pothos, E.: Primary Dissociated Midbrain Dopamine Cell Cultures from Rodent Neonates., J Vis Exp 21, 820, 2008
大鼠 Wistar大鼠, 4 天Hippocampal neuronsPDS kit: per instructionsNeurobasal AObradovic Darja, Gronemeyer Hinrich, Lutz Beat, Rein Theo: Cross-talk of vitamin D and glucocorticoids in hippocampal cells, J Neurochem 96, 500-9, 2006
大鼠SD大鼠, 新生的astrocytesPDS kit: per instructionsDMEMLacroix-Fralish, M., Tawfik, V., Nutile-McMenemy, N., Harris, B. and Deleo, J.: Differential Regulation of Neuregulin 1 Expression by Progesterone in Astrocytes and Neurons., Neuron Glia Biol 2, 227, 2006
大鼠大鼠, 2 月Spinal cord progenitor cells PDS kit: see referenceNeurobasal AMothe, A., Kulbatski, I., Van Bendegem, R., Lee, L., Kobayashi, E., Keating, A., and Tator, C.: Analysis of Green Fluorescent Protein Expression in Transgenic Rats for Tracking Transplanted Neural Stem/Progenitor Cells, J Histochem Cytochem 53(10), 1215, 2005
Hybrid toxins
大鼠SD大鼠, E19Dorsal root gangliaPDS kit: per instructionsMEM/Ham's F12Gavva NR, Tamir R, Qu Y, Klionsky L, Zhang TJ, Immke D, Wang J, Zhu D, Vanderah TW, Porreca F, Doherty EM, Norman MH, Wild KD, Bannon AW, Louis JC, Treanor JJ.: AMG 9810 [(E)-3-(4-t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4] dioxin-6-yl)acrylamide], a novel vanilloid receptor 1 (TRPV1) antagonist with antihyperalgesic properties, J Pharmacol Exp Ther 313, 474, 2005
大鼠 Wistar大鼠, E15Vomeronasal receptor neuronsCollagenase/Dispase: 0.1%DMEM/F12Moriya-Ito K, Osada T, Ishimatsu Y, Muramoto K, Kobayashi T, Ichikawa M: Maturation of vomeronasal receptor neurons in vitro by coculture with accessory olfactory bulb neurons, Chem Senses 30, 111, 2005
Papain: 0.5 u/ml
大鼠SD大鼠, 1-2 天Cortical astrocytesPapain: see referenceDMEMFloyd Candace L, Gorin Fredric A, Lyeth Bruce G: Mechanical strain injury increases intracellular sodium and reverses Na+/Ca2+ exchange in cortical astrocytes, Glia 51, 35-46, 2005
大鼠SD大鼠, 0-2 天Superior cervical ganglionCollagenase Type 4: 20 u/mlDMEMPedraza Carlos E, Podlesniy Petar, Vidal Noemi?, Arevalo Juan Carlos, Lee Ramee, Hempstead Barbara, Ferrer Isidre, Iglesias Montse, Espinet Carme: Pro-NGF isolated from the human brain affected by Alzheimer's disease induces neuronal apoptosis mediated by p75NTR, Am J Pathol 166, 533-43, 2005
Trypsin: 0.25%
大鼠SD大鼠,雄性, 5-8 周Trigeminal neuronsPapain: 20 u/mlCMF HanksConnor, M, Naves, L. and McCleskey, E.: Contrasting Phenotypes of Putative Proprioceptive and Nociceptive Trigeminal Neurons Innervating Jaw Muscle in Rat., Mol Pain 1, 31, 2005
Collagenase: 0.3%
大鼠大鼠, 胎兒Brainstem and cortical neuronsPDS kit: per instructionsDMEMLovshin JA, Huang Q, Seaberg R, Brubaker PL, Drucker DJ:Extrahypothalamic expression of the glucagon-like peptide-2 receptor is coupled to reduction of glutamate-induced cell death in cultured hippocampal cells, Endocrinology 145, 3495, 2004
大鼠SD大鼠,雄性 150-220 克Dorsal root ganglion neuronsCollagenase Type 4: 0.125%DMEM/Ham's F12Hu Hong-Zhen, Gu Qihai, Wang Chunbo, Colton Craig K, Tang Jisen, Kinoshita-Kawada Mariko, Lee Lu-Yuan, Wood Jackie D, Zhu Michael X:2-aminoethoxydiphenyl borate is a common activator of TRPV1, TRPV2, and TRPV3, J Biol Chem 279, 35741-8, 2004
Trypsin: 0.05%
大鼠大鼠, 15 周NeuronsPapain: 0.2%Hibernate AEvans J, Sumners C, Moore J, Huenman MJ, Deng J, Gelband CH, and Shaw G: Characterization of mitotic neurons derived from adult rat hypothalamus and brain stem, J Neurophysiol 87, 1076, 2002
大鼠SD大鼠,雄性, 270-330 克Spinal progenitor cellsPDS kit: with modificationsNeurobasal mediumLin CR, Wu PC, Shih HC, Cheng JT, Lu CY, Chou AK, Yang LC.:Intrathecal spinal progenitor cell transplantation for the treatment of neuropathic pain, Cell Transplant 11, 17, 2002
大鼠大鼠, 食魚貂, 8-9 周Adult progenitorPapain: 2.5 u/mlDMEM/F-12Lie DC, Dziewczapolski G, Willhoite AR, Kaspar BK, Shults CW, Gage FH.: The adult substantia nigra contains progenitor cells with neurogenic potential, J Neurosci 22, 6639, 2002
Deoxyribonuclease I: 250 u/ml
Neutral Protease: 1 u/ml
大鼠SD大鼠, 胚胎Sciatic Nerve and Gut Neural Crest Stem CellsCollagenase Type 4: 0.025%HBSSBixby, S., Kruger, G., Mosher, J., Joseph, N. and Morrison, S.: Cell-Intrinsic Differences Between Stem Cells from Different Regions of the Peripheral Nervous System Regulate the Generation of Neural Diversity.,Neuron 35, 643, 2002
Trypsin: 0.005%
Deoxyribonuclease I: 0.05%
大鼠大鼠Neurons, hippocampalPapain: 15 - 20 u/mlEagle's MEMLiu, Q., Kawai, H., and Berg, D.: B-Amyloid Peptide Blocks the Response ofa7-Containing Nicotinic Receptors on Hippocampal Neurons, PNAS 98 (8), 4734, 2001
(see reference)
大鼠大鼠, 新生的, 7 天 大NeuronsTrypsin: 0.25%MEM10Acosta, C., Fabrega, A., Masco, D., and Lopez, H.: A Sensory Neuron Subpopulation with Unique Sequential Survival Dependence on Nerve Growth Factor and Basic Fibroblast Growth Factor during Development, J Neurosci 21 (22), 8873, 2001
大鼠大鼠, 胚胎18-19 天Hippocampal neuronsPDS kit: see referenceDMEMMabuchi T, Kitagawa K, Kuwabara K, Takasawa K, Ohtsuki T, Xia Z, Storm D, Yanagihara T, Hori M, Matsumoto M: Phosphorylation of cAMP response element-binding protein in hippocampal neurons as a protective response after exposure to glutamate in vitro and ischemia in vivo, J Neurosci 21, 9204-13, 2001
大鼠大鼠, 胚胎, 天 18CorticalPapain: Neurobasal medium and DMEMO'Connor, S., Andreadis, J., Shaffer, K., Ma, W., Pancrazio, J., and Stenger, D.: Immobilization of Neural Cells in Three-Dimensional Matrices for Biosensor Applications, Biosensors & Bioelectronics 14, 871, 2000
大鼠大鼠, Wister, postnatal, P1-3 天Neurons, hippocampalPapain: 20 u/mlEBSSNeuhoff, H., Roeper, J., Schweizer, M.: Activity-Dependent Formation of Perforated Synapses in Cultured Hippocampal Neurons, Eur J Neurosci 11, 4241, 1999
大鼠SD大鼠, 雌性, pre克nantSciatic nervesTrypsin: 0.025%L-15 mediumMorrison, S., White, P., Zock, C., and Anderson, D.: Prospective Identification, Isolation by Flow Cytometry, and In Vivo Self-Renewal of Multipotent Mammalian Neural Crest Stem Cells, Cell 96, 737, 1999
Collagenase Type 3: 0.1%(see reference)
大鼠大鼠Spinal cordTrypsin: 0.133%HBSS and PIPESJohansson, C., Momma, S., Clarke, D., Risling, M., Lendahl, U., and Frisen, J: Identification of a Neural Stem Cell in the Adult Mammalian Central Nervous System, Cell 96, 25, 1999
(包括 小鼠)
大鼠SD大鼠, 胚胎Sciatic nervesTrypsin: 0.025%L-15 mediumMorrison, S., White, P., Zock, C., and Anderson, D.: Prospective Identification, Isolation by Flow Cytometry, and In Vivo Self-Renewal of Multipotent Mammalian Neural Crest Stem Cells, Cell 96, 737, 1999
Collagenase Type 3: 0.1%(see reference)
大鼠大鼠, 小白鼠), 2-5 天 大Neurons, hippocampalPapain: 20 u/mlEBSSWilding, T., and Huettner, J.: Activation and Desensitization of Hippocampal Kainate Receptors, J Neurosci 17 (8), 2713, 1997
大鼠SD大鼠和Wistar大鼠, 新生的, 0-21 天, 兩者之間任意一個Myenteric gangliaTrypsin: 0.05%MEM-HEPESSchafer, K., Saffrey, M., Burnstock, G., and Mastres-Ventura, P.: A New Method for the Isolation of Myenteric Plexus from the Newborn Rat Gastrointestinal Tract, Brain Res Proto 1, 109, 1997
大鼠大鼠, 幼崽, 24-48 小時大Neurons, hippocampalPapain: 20 u/mlHarvest bufferHall, R., and Soderling, T.: Differential Surface Expression and Phosphorylation of the N-Methyl-D-Aspartate Receptor NR1 and NR2 in Cultured Hippocampal Neurons, J Biol Chem 272 (7), 4135, 1997
大鼠SD大鼠, 成年的, 250-300克, P8Neurons, DRGNeutral Protease: 0.5%L-15 w/ CO2Davies, S., Fitch, M., Memberg, S., Hall, A, Raisman, G., and Silver, J.:Regeneration of Adult Axons in White Matter Tracts of the Central Nervous System, Nature 390, 680, 1997
大鼠大鼠, 1-4 天 大Neurons, hippocampalPapain: 20 u/mlMEMTwitchell, W., Brown, S., and Mackie, K.: Cannabinoids Inhibit N- and P/Q-Type Calcium Channels in Cultured Rat Hippocampal Neurons, J Neurophysiol 78, 43, 1997
大鼠SD大鼠, 食魚貂Hippocampal neuronsPapain: 0.2%HibernateA/B27Brewer, G.J.: Isolation and culture of adult rat hippocampal neurons of any age, J Neurosci Methods 71, 143, 1997
大鼠大鼠, E18Hippocampal neuronsPapain: 20 u/mlMEMLiu QY, Schaffner AE, Li YX, Dunlap V, Barker JL: Upregulation of GABAA current by astrocytes in cultured 胚胎rat hippocampal neurons, J Neurosci 16, 2912-23, 1996
Deoxyribonuclease I: 0.01%
大鼠大鼠, 1-6 天DRG neuronsPapain: 20 u/mlHBSSRobertson, S., Rae, M., Rowan, E. and Kennedy, C.: Characterization of a P2X-Purinoceptor in Cultured Neurones of the Rat Dorsal Root Ganglia., Br J Pharmacol 118, 951, 1996
Collagenase Type 1: 150 u/ml
Neutral Protease: 0.8%
大鼠大鼠, 幼崽, 14 天 大Neurons, sympatheticNeutral Protease: 0.24%HBSSMcFarlane, S., and Cooper, E.: Extrinsic Factors Influence the Expression of Voltage-Gated K Currents on Neonatal Rat Sympathetic Neurons, J Neurosci 13 (6), 2591, 1993
大鼠,SD大鼠, 11-14 天 大, 29-32 克Basal forebrain neuronsTrypsin: 0.125%Gey's BSSAllen, T., Sim, J., and Brown, D.: The Whole-Cell Calcium Current in Acuy Dissociated Magnocellular Cholinergic Basal Forebrain Neurones of the Rat,J Physiol 460, 91, 1993
大鼠Hanover Wistar大鼠, 7-12 天Postnatal dopamine neuronsTrypsin: 0.035%(see reference)Rayport, S., Sulzer, D., Shi, W., Sawasdikosol, S., Monaco, J., Batson, D., and Rajendran, G.: Identified Postnatal Mesolimbic Dopamine Neurons in Culture: Morphology and Electrophysiology, J Neurosci 12 (11), 4264, 1992
大鼠SD大鼠, 18-天-大, 胎兒HippocampalTrypsin: 0.2%HBSSCheng, B., and Mattson, M.: IGF-I and IGF-II Protect Cultured Hippocampal and Septal Neurons Against Calcium-Mediated Hypoglycemic Damage, J Neurosci 12 (4), 1558, 1992
大鼠大鼠Stem, neural crestCollagenase: 0.075%Ringer's solutionStemple, D., and Anderson, D.: Isolation of a Stem Cell for Neurons and Glia from the Mammalian Neural Crest, Cell 71, 973, 1992
大鼠SD大鼠, 懷孕的, 查爾斯河Pyramidal neuronsTrypsin: 0.027%HEPESBuchhalter, J., and Dichter, M.: Electrophysiological Comparison of Pyramidal and Slate Nonpyramidal Neurons in Dissociated Cell Culture of Rat Hippocampus, Brain Res Bull 26, 333, 1991
Nonpyramidal neurons
大鼠SD大鼠, 胚胎, 19-21 天 妊娠期Schwann, dorsal root gangliaTrypsin: 0.25%HBSS, CMFMithen, F., Reiker, M., and Birchem, R.: Effects of Ethanol on Rat Schwann Cell Proliferation and Myelination in Culture, In Vitro Cell Dev Biol 26, 129, 1990
大鼠大鼠 HeartTrypsin: 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
(包括 牛)Adrenal chromaffin 
 Paraneurons
大鼠SD大鼠, 8 天 大 幼崽Cerebellar neuronsTrypsin: 0.025%Eagle's MEMNovelli, A., Reilly, J., Lysko, P., and Henneberry, R.: Glutamate Becomes Neurotoxic Via the N-methyl-D-aspartate Receptor When Intracellular Energy Levels are Reduced, Brain Res 451, 205, 1988
大鼠大鼠,胎兒, 18 天 大Hippocampal neuronsTrypsin: 0.2%Eagle's MEMMattson, M., and Kater, S.: Isolated Hippocampal Neurons in Cryopreserved Long-Term Cultures: Development of Neuroarchitecture and Sensitivity to Nmda, Int J Dev Neurosci 6 (5), 439, 1988
大鼠大鼠, 產(chǎn)后Septal neuronsPapain: 0.05%PBS, CMFHatanaka,H., Tsukui, H., Nihonmatsu, I.: Septal Cholinergic Neurons From Postnatal Rat Can Survive In The Dissociate Culture Conditions In The Presence Of Nerve Growth Factor, Neurosci Lett 79, 85, 1987
大鼠大鼠, 小白鼠, 1-15 天Neurons, visual cortexPapain: 20 u/mlBSSHuettner, J., and Baughman, R.: Primary Culture of Identified Neurons From the Visual Cortex of Postnatal Rats, J Neurosci 6, 3044, 1986
(see reference)
大鼠SD大鼠, 雌性, 4 月 大CNS cellsTrypsin: 0.25%EBSSWood, P., and Bunge, R.: Evidence That Axons are Mitogenic for Oligodendrocytes Isolated From Adult Animals, Nature 320, 756, 1986
大鼠大鼠, 胚胎, 15 天Dorsal horn neuronsTrypsin: 0.025%Ham's F-12Jahr, C. and Jessell, T.: Synaptic Transmission between Dorsal Root Ganglion and Dorsal Horn Neurons in Culture: Antagonism of Monosynaptic Excitatory Postsynaptic Potentials and Glutamate Excitation by Kynurenate,J Neurosci 5, 2281, 1985
Spinal
大鼠大鼠, 胎兒, 18-20 天Hippocampal neuronsTrypsin: 0.25%HBSS, CMFBartlett, 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, 產(chǎn)后Ganglion, retinaPapain: 12.5 u/mlHBSS w/5 mM HEPESLeifer, D., Lipton, S., Barnstable, C., and Masland, R.: Monoclonal Antibody to Thy-1 Enhances Regeneration of Processes by Rat Retinal Ganglion Cells in Culture, Science 224, 303, 1984
大鼠SD大鼠, 幼崽RetinaTrypsin: 0.25%Ham's F-12Sarthy PV, Curtis BM, and Catterall WA.: Retrograde Labeling, Enrichment, and Characerization of Retinal Ganglion Cells from the Neonatal Rat, J Neurosci 3 (12), 2532, 1983
大鼠大鼠, W/FU, 5-8 天Neurons and glialTrypsin: 0.25%MEMRaff, M., Fields, K., Hakomori, S., Mirsky, R., Pruss, R., and Winter, J.:Cell-Type-Specific Markers for Distinguishing and Studying Neurons And the Major Classes of Glial Cells in Culture, Brain Res 174, 283, 1979
(see reference)
大鼠大鼠, 胎兒Neurons, sympatheticTrypsin: 0.25%L-15 or HBSS, CMFWakshull, E., Johnson, M., Burton, H.: Postnatal Rat Sympathetic Neurons In Culture. 1. A Comparison With 胚胎Neurons, J Neurophysiol 42, 1410, 1979
大鼠 Wistar/Furth大鼠, 新生的SchwannTrypsin: 0.25%DMEMBrockes, J., Fields, K., and Raff, M.: Studies on Cultured Rat Schwann Cells. I. Establishment of Purified Populations From Cultures of Peripheral Nerve,Brain Res 165, 105, 1979
(包括 牛)
大鼠大鼠, 胚胎Neurons, corticalTrypsin: 0.027%MEMDichter, M.: Rat Cortical Neurons in Cell Culture: Culture Methods, Cell Morphology, Electrophysiology, and Synapse Formation, Brain Res 149, 279, 1978
大鼠大鼠, 新生的Neurons, sympatheticCollagenase Type 1: 0.01%Hank's solution, CFReichardt, L., Patterson, P.: Neurotransmitter Synthesis and Uptake by Isolated Sympathetic Neurones in Microcultures, Nature 270, 147, 1977
大鼠大鼠, 新生的Neurons, superior cervical gangliaTrypsin: 0.1%Basal L-15 mediumMains, R., and Patterson, P.: Primary Cultures of Dissociated Sympathetic Neurons I. Establishment of Long-Term Growth in Culture and Studies of Differentiated Properties, J Cell Biol 59, 329, 1973
蠑螈蠑螈, 18-25 cmRetinaPapain: 14 u/mlSalineTownes-Anderson, E., MacLeish, P., and Raviola, E.: Rod Cells Dissociated from Mature 蠑螈 Retina: Ultrastruture and Uptake of Horseradish Peroxidase, J Cell Biol 100, 175, 1985
蠑螈蠑螈 (A.tigrinum)Photoreceptors, retinaPapain: 0.05%(see reference)Bader, C., MacLeish, P., and Schwartz, E.: Responses to Light of Solitary Rod Photoreceptors Isolated From Tiger 蠑螈 Retina, Proc Natl Acad Sci U S A 75, 3507, 1978
貝類動物海兔NeuronsNeutral Protease: 1.0%L-15-ASWLee, A., Decourt, B. and Suter, D.: Neuronal Cell Cultures from Aplysia for High-Resolution Imaging of Growth Cones., J Vis Exp 12, 662, 2008
貝類動物蝸牛(旋節(jié)螺), 白化病的, 成年的Buccal ganglia; SLT muscleTrypsin: 0.2%DMEMZoran, M., Doyle, R. and Haydon, P.: Target Contact Regulates the Calcium Responsiveness of the Secretory Machinery During Synaptogenesis,Neuron 6, 145, 1991
貝類動物田螺(旋節(jié)螺) 白化病的, 成年的Somata, buccal gangliaTrypsin: 0.2%Antibiotic saline, Leibowitz 50%Haydon, P.: The Formation of Chemical Synapses Between Cell-Cultured Neuronal Somata, J Neurosci 8, 1032, 1988
貝類動物旋節(jié)螺Buccal gangliaTrypsin: 0.2%L-15 mediumCohan, C., Haydon, P., and Kater, S.: Single Channel Activity Differs in Growing and Nongrowing Growth Cones of Isolated Identified Neurons of Helisoma, J Neurosci Res 13, 285, 1985
貝類動物軟體動物, 青年的 1-2 克 或者成年的50-100 克NeuronsProtease: 1%L15 mediumSchacher, S., and Proshanksy, E.: Neurite Regeneration by Aplysia Neurons in Dissociated Cell Culture: Modulation by Aplysia Hemolymph and the Presence of the Initial Axonal Segment, J Neurosci 3 (12), 2403, 1983
LUQ cells
貝類動物海兔LUQ cellsProtease: 1%L15 mediumCamardo, J., Proshansky, E., and Schacher, S.: Identified Aplysia Neurons Form Specific Chemical Synapses in Culture, J Neurosci 3 (12), 2614, 1983
RUQ cells
龜 (紅耳龜)RetinalPapain: 0.1% (13.5 u/mg)Kreb's RingerLam, D.: Biosynthesis of Acetylcholine in Turtle Photoreceptors, Proc Natl Acad Sci U S A 69, 1987, 1972

 

我們公司zui大優(yōu)勢是強大的采購,

1:基本什么都能進口,血清,抗體,耗材,還有部分限制進口的,

2:貨品全,現(xiàn)經(jīng)營過700多個品牌,基本所有生物試劑耗材都可以進口,特別是冷偏的產(chǎn)品那就更有優(yōu)勢,

3:提供加急服務,一般1-2周到貨,超過時限加急費全免

4:價格公道,絕大部分價格有優(yōu)勢,當然不能保證100%產(chǎn)品都是,因為意味著沒有服務.

5:良好的信譽,大部分客戶我們提供貨到付款服務,客戶包括清華,北大 交大 復旦,中山等100多所大學,ROCHE,阿斯利康,國藥 ,fisher等500多家公司

6:我們還是Santa,Advanced Biotechnologies Inc, Worthington Biochemical ,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. Dako,Inc, GePromegaridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,Worthington Biochemical , Polysciences,prospecbi, QA-BIO,quickzyme,FULLERLABS,INC,sterlitech;sysy,TriLink BioTechnologies,Inc;worthington-biochem,zyagen等幾十家國外公司授權(quán)代理。
7:我們還是invitrogen,qiagen,Worthington Biochemical am,sigma;Promega,roche,merck, rnd,BD, GE,pierce,BioLegend等*批發(fā),歡迎合作。

欧美美女视频| 成人在线观看一区| 九九色欲网| 色综合丁香婷婷| 色九网| 久久这里只精品| 大地资源色婷婷视频在线| 亚洲人成www在线播放| 超碰在线免费观看日韩| 久久狠婷婷| 99久久婷| 激情色情五月天| 五月婷婷在线观看黄| 婷婷射图| 涩 五月 婷婷 狠狠| 婷婷五月天堂| 激情综合五| 久热免费视频| 成人 九九九九| 久久精品4| 天天色综合网吨吧| 午夜婷婷| 丁香五月ⅤA久久久| 综合激情婷婷| 天天摸天天日天天舔| GOGOGO免费高清日本TV| 青青青国产手线观看视频2019| 39视频第二区| 五月丁香六月婷婷开心网| 91视频综合网| 婷婷伊人綜合中文字幕| 激情久久综合| 色噜噜狠噜噜视频| 99资源人人| 人人人舔人人人操人人人摸人人人97| 五月婷婷五月天| 久久久ww| 激情丁香五月天图片| 欧洲毛片基地c区| 色六月 婷婷| 超碰超碰在线| 日韩操人| 性热视频99精品| 婷婷六月久久| A久久| 在线资源av-超碰中文在线-成人AV| 婷婷六月开心网| 色情丁香五月婷婷精品| 亚洲六月色婷婷| 婷婷五亚洲| 亚洲人成www在线播放| 成人五月天在线观看| 激情婷婷六月天| 九九综合色| 婷婷五月天影院| 婷婷丁香人妻天天| 国产亚洲成人综合| 天天舔天天摸视频| 91女人18毛片水多国产| 五月香蕉婷婷| 99热在线看| 久久九九99| 天天爽夜夜操| 五月停停丁香| 亚洲丁香五月天在线视频| 五月婷久久| 色五月激情综合| 六月丁香婷| 激情综合网五月天天| 91无码高清| 色欧美日| 91无码视频| 五月丁香免费视频| 亚洲经典三级| 日韩小视频在线99| 九九人人操| 五月丁香婷婷激情在线| 精品水蜜桃久久久久久久| 激情深愛五月視頻| 夜夜躁爽日日| 丁香六月婷婷一区| 激情小说五月天| 开心五月激情五月丁香五月婷婷| 成人看片网站| 深爱激情婷| 色吧婷婷五月亚洲| 99精品综合视频| 综合九九中文字幕| 青草青草视频2免费观看| 超级碰 久久9| 狠狠色噜噜狠狠狠888| 国产44页| 久久丁香五月天| 五月天无码视屏播放| 超碰狠狠干99| 四色AVwww| 国内自拍97在线| 久久综合网免费视频| 色爱综合网| 婷婷色日本| 91超级碰人人操| 色色日本欧美| 色色色五月婷婷| 麻豆AV一区二区三区| 色综合激情| 日本欧美成人片AAAA| 精品视频这里只有精品| 拍色综合| 99热国产这里只有精品| 婷婷久久久久久久| 丁香综合久久| 午夜天堂一区人妻| 深爱激情av| 天天综合网~91| 激情小说视频图片| 俺也去在线久久精品23欧美综合视频网站,丰满人妻一区二区三区在线视频53,丰满 | 999激情视频| www.久久久久久| 婷婷色色五月天| 丁香五月婷婷成人综合| 五月天婷婷Av| 色色a| 激情五月天伊人av| 五月婷婷综合久久| 色级停停| 99ri视频在线播放| www,五月丁,com| 日韩av高清| 伊人激情影院| 五月综合在线婷婷图片| 亚洲天堂九九九| 色播丁香| 99热都是精品| 亭亭色天香| 婷婷丁香五月激情密臀av| 91黄址| 中文字幕丰满乱孑伦无码专区| 蜜桃精品免费久久久久影院| 日本视频99| 色婷綜合网| 踪合专区啪啪| 丁香五月婷婷五月基地| 色狠狠色噜噜AV天堂五区| 五月色丁香视频精品| 99热91| 五月婷婷丁香六月| 婷婷丁香五月激情| 99热在线这里| 日本成人噜噜噜噜噜| 9月色婷婷| 精品人妻一区二区三区四区不卡在| 久久黄色片| anquye伊人| 色很久综合| 久久女婷| 婷婷综合五月天亚洲综合| 九九视频免费| 91丨九色丨白浆秘| 中文成人在线| 97日本在线播放| 九九精品亚洲| 激情五月天在线视频| 日本色婷婷五月天成人电影| 亚洲激情网| 色yeye色综合| 五月婷婷久久网| 五月色综合| 日韩在线观看亚洲| 五月天婷婷综合网| 亚洲视频99| 操操碰| www久久久久久| 欧州婷婷五月天综合| 高清无码入口| 五月婷婷AV| 无套内谢少妇毛片A片小说| 99re在线这里只有精品视频首页| 丁香六月啪| www,超碰| 9超碰在线| 91精品91久久久久77777| 五月丁香六月| 五月激情影视| 欧美性爱五月天| 婷婷五月综合亚洲| 五月四色色| 婷色五月天| 久久九九思思| 精品水蜜桃久久久久久久| 99久久99久久综合| 五月婷婷激情久久| 日本欧美成人片AAAA| 色婷婷综合久久久久| 99热国产在线| 免费视频这里只有精品| 一级操逼内射在线视频| 五月色导航| 成人va在线观看视频| 五月丁香婷婷啪啪综合| 欧美激情伊人| 在线你懂的亚洲欧| 男人視頻站| 丁香五月激情啪啪啪| 狠狠色综合五月人人| 激情欧美日韩一区二区| 婷婷五月在线观看| 九九视频在线观看视频6 | 五月丁香激情综合啪啪| 97精品欧美91久久久久久久| 91日韩在线| 亚洲色在线观看| 色婷婷影院| 欧美成人A片AAA片在线播放 | 国产无人区大片| 丁香五月色五月| www.深爱激情| 另类激情五月在线视频欧美| 国产婷婷五月色情综合| 五月天成人在线播放| 日本狠狠干| 中文字幕成人影视| 日日夜夜天天综合| 一区三区三区不卡| 亚洲中文字幕翔田千里| 日本黄色精品| 色婷婷大香蕉| 亚洲中文字幕AV| 婷婷六月激情小说网| 九九大香视频| 五月天婷婷六月激情网| 五月丁香六月婷婷玖玖| 五月丁香啪啪伦理电影| 熟女网站久久| 国产成人精品一区二三区熟女在线 | 亚洲欧洲中文日韩久久AV乱码| 日韩中文字幕| 婷婷伊人综合| 欧洲一区二区| 天天摸,天天爽| 日日狠夜夜狠| 猛烈顶弄H禁欲老师H春潮| 黄网免费观看| 不卡的无码高清的免费| 97操男人的天堂| 性色婷婷| 婷婷国产日本欧美| 狠狠操狠狠操AV| 九九精品在线观看视频6| 开心五月四房播播| 欧美大片免费播放器| 日本二级毛片二级毛片| 欧美超级视频97| 女性自慰系列第五页| 婷婷五月开心中文字幕色| 九九在线这里只有精品视频| 二色AV| 99久久精品免费看国产一区二区 | 欧美人妻一区二区| 婷婷香五月天| 色激情网| 丁香五月情色| 婷婷色五月天在线| 五月天精品| 90色免费视频| www狠狠| 色情久久久| 99热这里| 99热啪啪| 97色永久免费视频| www.五月天。com| 猴哥影院免费看电影| 囯产精品久久欠久久久久久九大| 99视频啪啪| 婷婷激情四射| 五月丁香婷婷成人网| 久久538| 久热精彩视频98| 婷色五月| 婷婷五月激情六月| 久热9| 婷婷在线日韩综合| 狠狠色狠狠干| 丁香花在线电影小说观看| 免费无码毛片一区二区A片| 1024你懂的欧美曰韩| 五月天激情站| 香蕉视频91| 成人草榴视频| 日韩色色小视频| 亭亭玉月丁香| 爱操天堂| 亚洲天天免费| 337p大胆噜噜噜噜噜91Av| www.金莲av| 99热福利| 五月婷婷丁香网| 久草热在线视频| 色五月婷婷天堂| 五月亭亭激情综合| 久色视频| 亚洲婷婷91丁香| 婷婷综合五月天亚洲综合| 中文字幕无码人妻少妇免费视频| 色色色色色综合| 六月婷婷狠狠做| 97精品综合| 色吊丝av中文字幕| 国产色五月| 中文字幕有多少字| 99爱爱| 五月天色区| 激情婷婷丁香| 丁香五月六月婷婷自拍| 色情成人五月天| 久久婷婷五月综合激情国产| 五月天婷婷在线啪啪视频| 99色综合网| 99热在线网站| 天天操夜夜爽歪歪| 日逼AV影音先锋男人资源站| 高清无码 一区 二区 三区| 99操逼视频| 亚洲激情五月| 婷婷五月天第三页| 9色天堂| 天天檫天天爽| 在线播放人妻| 森林影视大全,最好看的2019年视频 | 26uuu欧美亚洲日韩| 六月婷婷五月丁香| 五月欧美色播| 狠狠色噜噜狠狠狠888| 棕合影院色色| 综合99综合久久久久久久| 欧美一线视频| 婷婷 丁香 精品| 天天色五月婷婷91久久久久久久| 色性五月天| 久久激情中文| 五月婷婷乱| 国产亚洲欧美日本一二三本道 | 超碰精品国产首页| 天天撸一撸| 99热在线看片| 五月天色视频| 婷婷五月丁香99| 婷婷六月色播| 69人人操人人爽| 影音先锋777xfplay色资源网站| 五月Huangsewang| 这里只有精品99www| 日日夜夜国产| 亚洲色综久久五月| 久久婷婷电影| 一级二级色大片| 久久久久久99精品无码| 国产 亚洲 在线| 超碰91在线| 激情五月丁香五月| 天天拍夜夜撸| 亚洲va成人va成人va在线观看| 婷婷中文字幕| 成人国产欧美大片一区| 九九精品视频在线6| 色欲久久99精品久久久久久| 婷婷狠狠干| 婷婷久久18| 久久9热| 久久九九爽| 六月婷婷开心| 99亚洲欧洲| 爱iii做iiii日| 99色免费观看全部| 97超碰在线免费观看| 久久久久久激情| 伊人影音无码一区二区三区 | 夜夜操少妇| 成人丁香婷婷| 在线视频reer6| 九九久久五月天| 免费视频无码| 五月综合影院| 亚洲黄3级片网站欧美| 91狠狠综合久久久| 五月综合激情综合久| 久久久潮喷-久久久九九-成人AV| 99啪99| 激情深爱综合| av操B网站| 无码激情AAAAA片-区区| 国产乱码久久| 色在线免费观看| 婷婷五月天综合AV| 日本色婷婷综合| 另类婷婷五月天啪帕帕| 人人综合久| 六月丁香视频网站| 中文字幕欧美精品久久| www超碰| 久久九九99字幕| 色色九区| a亚洲在线观看不卡高清| 丁香五月婷综合网| 亚洲精品国产成人AV在线| 亚洲视频一区| 辣椒视频| 99视频精品8| 97超级碰碰碰| 欧美午夜精品一区区电影| 丁香五月天激情免费在线观看AV777| 色五月色五天色情网| 亚洲色婷婷| 激情五月天影院| 人妻久久久久久久久妻久久久久久久久 | 亚洲色婷婷五月天| se色99| 午夜av网| 青青青国产最新视频在线观看| 久久爱婷婷| 99惹在线精品免费观看| 日hao1区| 超碰9799| 色999亚洲人成色| 丁香六月视频免费观看| 亚洲成人免费电影| 五月天色欧美| 一区二区视频在线观看高清视频在线 | 在线99精品| 亚洲欧洲中文日韩久久AV乱码| 免费观看大片视频 丁香婷婷 六月欧美| 夜夜爽天天| a在线免费v| 亚洲深喉aV| 色婷婷久久综合| 性天天中文网| 五月情丁香色| 九九热自拍| 视频一区二区在线| 中文字幕日产A片在线看| 亚洲顶级VA在线观看-高清完整版在线影院观看-S022AV | 国产一级片| 超碰人人妻| 无码人妻精品一区二区蜜桃色欲| 成人永久免费视频在线观看| 大陆肏屄视频| 丁香五月亚洲无码| 欧美婷婷| 成人丁香色| 5Www色5夜| 99久久人妻精品无码二区| 夜夜撸.com| 五月天久久综合婷婷丁香| 欧美激情 日韩无码 婷婷 五月天 久久婷婷丁香五月一二三 | 天天射夜夜骑| 91婷婷五月天综合视频| 国产1区2区3区在线观| 人橾人| 操草草草| 色五月婷婷成人| 亚洲爆乳无码精品AAA片蜜桃| 无码橾| 色婷五月天激情| 婷婷五月天视频在线观看| 婷婷五月天成人导航| 国产婷婷综合在线免费视频| ...婷婷五月综合不卡,国产在线手机| 激情五月四色| 婷婷情色开心五月天99| 青青青在线视频免费观看| 99热.com| 91热在线| 色色五月天网站| 天天干天天干天天| 天天日夜夜爽| www.五月天婷婷| 婷婷婷婷婷婷婷五月丁香| 五月涩涩网| 天天日人人爽| 色99在线观看| 深情六月婷婷综合久久| 婷丁香久综合| 九九激情网| 九九视频免费| 九九热视频精品2| 国产精品大香蕉| 99re视频在线播放| 97色婷婷成人综合在线观看| 99精品热视频| 日本69日人视频| 综合久久97| 91丨九色丨白浆秘| 久操大香蕉| 丁香花色色网| 99er6免费视频热播| 99久久99久久综合| 久久精品爱爱| 天天 日综合| 久久午夜理论| 伊人久久丁香婷婷六月五月综合| 狠狠插日日干撸| 婷婷五月天开心激情网| 五月天com| 特级毛片AAAAAA| 九九干视频| 五月婷婷久久久| se99视频| 免费看成人AA片无码视频吃奶| 亚洲高清自拍| 九月丁香网婷婷| 狠狠干狠狠操狠狠爱| 无码色综合| 亚洲旡码| 色五月丁香总合网| 激情婷婷六月| 蜘蛛女侠2003满天星免费观看| 免费亚洲婷婷| 五月婷婷av| 五月综合色| 91精品91久久久中77777| 激情AV综合| 丁香花婷婷五月天| 51XX午夜影福利| eeuss人妻| 九九一综合精品| 五月丁香婷婷成人综合网| 色噜噜狠狠色综合日日| 超碰人人操人人干| 亚洲乱码日产精品BD| 五月久久综合| 4399伦理午夜| 色色无码日韩| 337午夜福利| 五月激情小说| 五月婷婷在线视频| 九九成人电影婷婷| 黄网在线免费播放| 九九热只有精品6| 伊人久久五月天| 开心四房播播| 北京熟妇搡BBBB搡BBBB| 热久91| 综合婷婷久久| www.思思99热| 欧美 日韩 成人| 欧美肉大捧一进一出免费视频| 影音先锋五月婷婷| 亚洲另类婷婷综合| 国产精品久久久久久五月天加勒比 | 色六月丁香婷婷狠狠干| 久久激情五月婷婷| 色婷婷六月| 99久99久| 日本色色网| 五月丁香黄色| 九九热黄色| 亚洲综合五月天婷婷| 久久婷婷丁香| 五月天婷婷丁香视频| 免费无码又爽又刺激A片涩涩直播| 激情婷婷六月天| 超碰无码老师| 天天玩夜夜操天天爽| 亚艹艹| 亚洲欧美成人在线| 五月婷婷欧洲| 九月丁香婷婷综合激情| 激情五月婷婷在线观看| 久久人妻少妇嫩草AV| 日本欧特黄色刺激一区影视久精品无码| 这里只有精彩小视频视频网站| 色婷婷五月中文字幕在线dvd| 另类天堂| 丁香五月WWW| 欧美日韓成人亚洲精品另类| 五月天婷婷AV| 五月激香蕉网| 五月丁香激情综合啪| 天天操综合网| 精品成人在线| 婷婷五月激情的图片| 久久九九免费视频| 日韩一区二区在线免费观看| 色综合网址| 夜夜撸天天操| 99欧美| 色婷婷黄色网络| 色五月丁香婷婷久草| 色天堂婷婷| 99久热这里只有精品视频删减版| 激情六月天| 色婷婷色综合久久精品V| 综合激情伊人影视在线| 九九99热| 五月婷婷之综合激情在线| 婷婷成人综合免费视频| 97天堂| 激情综合色婷婷啪啪六月天| 色色cOm| 亚洲在线中文字幕2| 男女99免费视频| 亚洲无码色| 色域五月丁香| 色综合性视频| 亚洲V国产V欧美V久久久久久| 自拍偷窥99热| 久久久久久久久99精品| 欧美日韩日韩成人| 婷婷五月成人系列| 亚洲综合网在线| 国产精品五月丁香| 天天操综合网| 婷婷丁香五月综合| 日韩久久视频| 亚洲无码成人| 97色天堂| 久久9999| 特级毛片AAAAAA| 久久艹网| 久久激情五月婷婷| 久久九九99.www| 深爱激情69热| 色婷婷成人色网| 婷婷五月天亚洲图片| 婷婷伊人| 成人狠狠成人狠狠成人狠狠成人狠狠 | 激情五月综合| 婷婷综合久久| 99婷婷| 99日韩| 激情综合婷婷| 7777激情基地| 五月丁香啪啪综合| 亚洲婷婷五月天在线激情综合网| 久草婷婷| 色色色777| 四川BBB搡BBB搡多人乱亂| 亭亭玉立国色天香| 丁香五月天视频| 久久久久久久综合狠狠综合| www婷婷| 五月天天天操天天爽夜夜操| 亚洲精品V天堂中文字幕| 玖玖资源站国产| 成人做爰高潮A片免费视频| 成人五月天婷婷| 国产亚洲精品久久久久久郑州| 婷丁五月| 国产午夜亚洲精品一区| 天天操,天天插| 日本三级第一页| 久久99激情丁香婷婷小说网| 秋霞av不能| 五月小说| 婷婷综合激情| 国产露脸150部国语对白| 丁香五月综合图片在线观看| AV中文网| 永久精品| 伊人色综合影院视频| 色播五月丁香| 国产五月天激情小说| 美女婷婷激情亚洲| 韩国激情五月天综合网| 久久久久8888| 天天综合精品| 婷婷五月色丁香在线看| 另类图片五月天激情| 搡BBBB搡BBB搡| 亚洲精品无码久久| 99久久精品免费看国产一区二区| www.俺去也com| 久久免费试看120秒| 8区视频在线| 狠狠艹狠狠艹| 播丁香五月婷婷欧美| 99久久五月婷婷| www色色com| 欧美精品久久久久久久小说| 五月丁香婷爱在线| 超碰色色综合| 中文字幕精品无码一区二区| 亚洲激情97五月天| 婷婷丁香五月天影院 | 玖久精品视频9| 丁香六月青青草| 人妻中文在线| 九玖欧洲亚洲| 在线资源av-超碰中文在线-成人AV| 色婷婷丁香五月天| 婷婷六月天亚州| 亚洲AVDVD| 这里只有精彩视频| www.开心激情| 91se在线视频| 日韩艹比| 影音先锋天天日| 色久在| AV大片在线观看| 久操人妻| 色狠狠色狠狠| 99久在线观看| 亚洲三级无码| 色五月超碰| 森林影视大全,最好看的2019年视频 | 九一99| 婷婷精品视频| 大香蕉福利导航| 大香蕉五月天婷婷| 久久人妻在线| 99九九久久| 婷婷九月久久| 天天干夜夜想| 丁香五月Av| 丁香午月AV中文字幕| 极品少妇婷婷五月| 久久久无码精品成人A片小说 | 五月丁香久久网| 激情五月婷婷在线| 91超级碰碰| 涩玖玖免费视频| 亚洲视频一区| 免费的视频APP网站入口| 成人小说 五月天 婷婷| 97色欧美| 激情综合视频| 开心深爱激情网| 伊人五月综合网| 天天爽天天日人人爱 | 无码少妇高潮喷水A片免费| 97色色视频| 亚洲第精品| 九色婷婷| 色色五月综合| AV中文在线| 色色操| 2025天天日爽| 六月五月婷婷| 六月婷婷之青青草| 日本系列_4页_777FP| 97碰碰草| 国产在线aaa片一区二区99| 天天综合色丁香| 丁香五月在线看| 99热6这里只有精品6| 天天日夜夜欢| 久操大屁股女人av| 综合天堂AV久久久久久久| 色五月婷婷久久| 天堂在线中文| 99热国产这里只有精品| 99久久思思| 99热这里只有99| 中文精品在| 99啪啪骑| 久久婷婷五月综合色区| 国产暴力强伦轩1区二区小说| 91偷拍视频| 色五月婷婷一二| 久久这里只有国产视频| 亚洲激情高潮| 99热 这里只有精品 国产 日韩| a在线免费v| 激情五月婷色| 五月丁香亭亭电影久久| 久久99国产综合精品免费| 人人爱操| 在线婷婷| 99九九99九九九视频精彩| 五月婷婷综合丁香视频| 色五XX| 久久AV电影| 欧美综合婷婷网| 婷婷丁香五月天色播网站| 九月激情网| 啪啪99| 三级毛片7979| 97碰在线免费观看| 久久婷婷五| 色色97丁香婷婷五月天| 超碰婷婷五月| 99在线视频观看| 极品精品一区二区三区在线| 久久婷五月| AV亚洲AV永久无码精品网| 中文字幕免费高清电视剧| 天天玩夜夜操| 深爱五月婷婷| 天天爽天天操| 国产成人片| 四川BBB搡BBB搡多人乱亂| 色色色色网站| 狠狠干,狠狠操| 99热亚洲| 成人片黄网站色大片免费毛片| 人妻少妇色综合| 在线只有精品| 风流少妇A片一区二区蜜桃| 中国女人做爰A片| 538在线精品| 丰满少妇猛烈A片免费看观看| 日本欧美成人片AAAA| 精品一区二区三区四区五区六区 | 波多婷婷久久| 超碰A V在线| 免费看片在线观看| 国产黄色一级片| 激情爱爱网站超大免费| 另类A片| 婷婷亚洲日本| 久久杏爱视频| 日日操日日干| 国产色五月| 五月天婷婷激情综合| 少妇综合网| 人妻视频在线| 丁香九月婷婷| 亚洲V国产V欧美V久久久久久 | 五月天综合网| 婷婷五月电影院| 丁香五月手机在线| 92久久精品一区二区| 97色五月婷婷在线| 成片免费观看大全| 1024婷婷综合久久五月天| 激情视频网址| 国产成人VA| 亚洲免费99| 色色色色色网| 91精品久久久久| 成人精品视频99在线观看免费| 99热官网| 欧美六月婷婷| 99热在线精品播放| 婷婷色播色五月五色五月天色妇| 第六色在线| 99在线小视频| 91精品无码| 欧美日本免费一道免费视频| 丁香情色五月| 99色免费| 五月婷婷六月丁香| 久热这里精品免费| 五月天色婷婷综合| 丁香五月影院| AⅤ网站在线看| 色涩影院六月丁香| 一级片sese片.COM| 思思热久久婷婷五月天| 狠狠色丁香婷婷综合| 人人草人人爱| 国产亚洲AV人片在线| av激情在线| 日日噜狠狠色综| 79色色色色| 琪琪色五月天| 狠狠爱综合网| 日本人人草草| 色噜噜狠狠色综合日日免费| 综合五月婷婷| 色五月丁香五月| av操B网站| 日韩999| 日韩欧美骚货| 成人av在线电影| 亚欧洲乱码视频一二三区| 操操操91| 天天做综合| 五月色综合网| 五月天婷婷激情| 3DAV亚洲香蕉久久 一区二区| 五月色网| 九九色综合视频| 五月婷婷香| 大香蕉久久伊人网| 女性自慰系列第五页| www五月天com| 精品一二三区久久AAA片| 成人一区在线观看| 亚洲五月婷婷在线| 免费无码又爽又刺激A片涩涩直播| 欧美精品在线观看| 色天堂97| 五月色婷婷激情| 久久久网站| 色情成人五月天| 六月婷婷深深爱| 99爱在线视频| 好青青在线视频观看视频| 日本九九视频| 色情五月天A片| 99无码视频| 色播色丁香五月| 五月婷婷综合激情| 色婷天天| 激情电影五月婷婷| 精品九九在线观看| www999日韩精品| 国产AV成人精品| 伦乱美欧| 日韩在线视频网站| 大香蕉久久伊人网| 婷婷免费成人视频| 午夜成人网站在线观看| 婷婷9月天| 美臀自射自家人妻| 99自拍视频网站| 91熟妇大香蕉| 第六色在线| 色婷婷影院| 婷婷五月天中文字幕| 天天干天天射综合网| 色婷婷丁香五月| 在线观看亚洲视频影院| 色吧五月婷婷| 超碰97在线操| 激情五月深爱五月| 影音先锋一区| 亚洲av成人电影在线观看| 艾小青av| 成熟妇人A片免费看网站| 欧美在线97| 五月天伊人| 玖玖婷婷五月天毛片| 丁香五月婷婷色综合基地| 99无码视频| 久久99网址| www.婷婷五月| 久久丁香婷| 激情美女五月天激情在线| 欧美成人精品A片免费一区99| 在线视频你懂得| 天天肏高清在线| 91九色无码内射| 激情五月天色婷婷| 日日操夜夜擼| 天天日人人| 99精品国产在热久久| 激情AV| 超碰人人操人人干| 欧美天堂久久| 日日夜夜干| 色五月婷婷激情基地| 99这里只有精品|v| 激情电影五月婷婷| 丁香婷婷在线| 日韩成人中文| www,五月丁,com| 涩五月婷婷| 97色伦另类图片小说视频 | 婷婷五月骚厕所| 婷婷视频在线| 五月丁香六月婷婷a v| 少妇熟女视频一区二区三区| 色五月婷婷 成人| 狠狠操狠狠插| 丁香六月天AV| 色婷婷成人做爰A片免费看网站| 成人五月天婷婷| 色五月婷婷基地| 国产中文亚洲欧美日韩性交| 婷婷婷婷色| 丁香六月婷婷一区| 99网| 人色五月天婷婷| 97精品综合| 天天干天天干天天干天天干天天干天天干天天| 日韩一区二区三区免费视频| 五月综亚洲| 99操99| 五月丁香婷庭在线| 91国产精品视频播放| 丁香五月婷婷色情综合| 五月天色欧美| 偷吃高潮H闺蜜H宋冉| 台湾无码A片一区二区| 丁香激情久久| 开心五月婷婷五月| 嫩草AV久久伊人妇女超级A| 五月婷婷香蕉| www.91五月| 天天日,天天干,天天操| 欧美啪啪五月天| 99乱视频| 天天激情5月天亚洲| 777精品久无码人妻蜜桃| 婷婷区日本| seav天堂| 五月天网站免费欧美| 婷婷大乡焦噜噜| 丁香五月婷婷啪啪| 成人色站,在线视频,看片-SS1AV| 亚洲天堂免费看| 懂色AⅤ| 九九九九九九综合| 亚洲欧美成人在线| 六月婷婷视频| 久久44| 久久AV无码乱码A片无码波多| 999热在线视频| 国产成人精品一区二三区熟女在线| 国产精品-第3页-91JQ就要激情网91JQ5.JQJQ926.XYZ | 91碰免费视频| 超碰高清在线| 成人色五月天婷婷| 日韩一区二区三区免费视频| 噜噜久| 国产ava| 国产成人片| 中文在线视频久1| 婷婷涩涩五月天| 国产人妻777人伦精品HD| 色婷婷在线视频综合| 丁香婷婷婷五月综合色情| 91Chinese在线| 久久婷婷五月综合激情国产| 色婷婷色和| 99色天堂| 99re热久久| 大香AV| 五月天激情网站| 五月开心深深爱激情综合| 91精产品自偷自偷综合| 中文字幕日产A片在线看| 99视频在线精品免费观看2| 国産精品| 超碰9799| 久大香蕉| RenRenSe在线视频网站| 美欧成人视频| 99精品视频在线观看| 久草热视频在线观看| 成人在线精品| 91re色综合视频| 超碰人人艹| 综合五月天天天天天五月| 伊人婷婷五月天av| 国产亚洲精品AAAAAAA片| 婷婷五月丁香在线视频| 丁香婷婷激情综合五月激情| 欧美五月婷婷综合| 久久精品99| 五月丁香六月婷婷视频| 色色婷婷五月天| 日本色啪| 激情五月天婷婷丁香| 五月婷婷天| 无码九九| 狠狠色狠狠爱| 久久91精品国产91| 五月丁香六月婷婷久久肏| 色婷綜合网| 大香蕉九操| 伊人狼人干| 狠狠干五月天| 久久爱婷婷| 激情五月天啪啪| 一区二区乱码视频| 丁香五月天久久| 综合网天天| A久久| 丁香五月天欧美| 成人视屏在线观看| 婷婷日本在线| 少妇性BBB搡BBB爽爽爽视頻| 五月四房播播| wwwC0maV五月花| 丁香五月自拍| 婷婷五月天中文字幕| www.91av.com| 丁香婷婷社区| 超碰网站在线观看| 丁香婷在线| 丁香五月婷婷姐| 99热在线观看精品| 影音先锋AV男人站| 日本亚洲欧洲另类图片| 中文字幕黄色片| 天天色天天| 婷婷激情啪啪| 久久只有精品| 亚洲操逼片| 日韩大片艹艹| 91久女| 日日躁夜夜躁狠狠久久AV| 97精品欧美91久久久久久久| 五月婷婷真爱激情网| 久久99热精品a片在线观看| www.激情五月天。com| 桃色激情五月天| 九月婷婷激情久久| 超碰日韩人妻在线| 99精品偷自拍| 综合久久婷婷| 97色天堂| 色婷婷的五月天| 丁香五月天网友自拍啪啪啪视频| 色播五月丁香婷婷| 五月丁香日本一抹本| 日本三级中国三级99| 欧美天天搞| 加勒比久热| 人妻视频一区而且二区| 一区二区视频在线观看高清视频在线| 日韩啊啊啊| 69精品人人人人| 日日噜噜夜夜狠狠久久丁香六月| 99精品在线|