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worthington 眼睛组织分离指南

发布者:艾美捷科技    发布时间:2022-06-01     
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原代细胞分离中,针对眼睛组织涉及到视网膜内皮、脑内皮细胞、视网膜细胞和角膜上皮 、米勒细胞等等的解离通常会使用酶,如何选择合适的酶来完成对应脑组织的解离,以及原代细胞采集的常规操作, 艾美捷针对眼睛相关的组织分类解离分种属、不同的细胞、酶组合推荐,推荐浓度、培养基以及参考文献,详细汇总指南:

眼睛组织分离指南

本worthington 眼睛组织分离指南描述了标准实验室程序; 为建立细胞分离方案提供了一种合乎逻辑的实验方法; 并列出了许多组织特异性参考资料,以帮助开发有效的方法。更多其他组织的解离指南,点击查看:各组织分离指南

 

眼睛
种属种属详情细胞名称酶组合推荐,含推荐浓度培养基参考文献
Bovine视网膜内皮 / Retinal endothelialCollagenase Type 4: 0.05%;
Deoxyribonuclease I: 0.02%;
Pronase: 0.02%
DMEMTerlizzi, V., Kolibabka, M., Burgess, J., Hammes, H. and Harmsen, M.: The Pericytic Phenotype of Adipose Tissue-Derived Stromal Cells Is Promoted by NOTCH2., Stem Cells 36, 240-251, 2018
Bovine周细胞 / PericyteCollagenase: 0.2%DMEMBryan, B. and D'Amore, P.: Pericyte Isolation and USe is Endothelial/Pericyte Coculture Models, Meth Enzymol 443, 315, 2008
Bovine微血管内皮 / Microvascular endothelialCollagenase/Dispase: 0.1%MEMBowman, P., Betz, A., and Goldstein, G.: Primary Culture of Microvascular Endothelial Cells From Bovine Retina, In Vitro 18 (7), 626, 1982
Chick, embryo, 6 day视网膜细胞 / Retinal cellsTrypsin: 0.005%;
Deoxyribonuclease I: 0.005%
DMEM/F12Jacob Vanessa, Rothermel Andrae, Wolf Peter, Layer Paul G: Rhodopsin, violet and blue opsin expressions in the chick are highly dependent on tissue and serum conditions, Cell Death Differ 180, 159-68, 2005
Chick embryo角膜上皮 / Corneal epitheliaCollagenase: 0.08%;
Trypsin: 0.08%
HBSSReenstra Wende R, Orlow Daniel L, Svoboda Kathy K H: ECM-stimulated signaling and actin reorganization in embryonic corneal epithelia are Rho dependent, Inv Ophthal Visual Sci 43, 3181-9, 2002
Chick embryo (also rat)视网膜 / RetinalTrypsin: 0.6%DMEMSeigel, G.: The Golden Age of Retinal Cell Culture, Mol Vis 5, 4, 1999
Chick, embryo, 10-14 day平坦,视网膜 / Flat, retinaTrypsin: 0.1%Tyrode's solution, CMFMoyer, M., Bullrich, F., and Sheffield, J.: Emergence of Flat Cells From Glia in Stationary Cultures of Embryonic Chick Neural Retina, In Vitro Cell Dev Biol 26, 1073, 1990
Fish, zebrafish视网膜 / RetinalPapain: 10 u/mlL-15Diekmann, H., Kalbhen, P. and Fischer, D.: Characterization of Optic Nerve Regeneration using Transgenic Zebrafish., Front Cell Neurosci 9, 118, 2015
Goldfish视网膜双极 / Retinal BipolarPapain: 30 u/mlHEPESGraffe, M., Zenisek, D. and Taraska, J.: A Marginal Band of Microtubules Transports and Organizes Mitochondria in Retinal Bipolar Synaptic Terminals., J Gen Physiol 146, 109-17, 2015
Human角膜基质细胞角质细胞 / Corneal stromal keratocytesCollagenase Type 1: 0.1%DMEM/F-12Yusoff,B, Riau, A., Yam, G., Halim, N. and Mehta, J.: Isolation and Propagation of Human Corneal Stromal Keratocytes for Tissue Engineering and Cell Therapy.,Cells Vol. 11,, , , 2022
Human米勒细胞 / Muller gliaPDS kit: per instructionsDMEMXu, N., Chen, Y., Dean, K., Lu, X., Liu, X., Wang, W., Dean, D., Kaplan, H., Gao, L., Dong, F. and Liu, Y.: Sphere-Induced Rejuvenation of Swine and Human Muller Glia Is Primarily Caused by Telomere Elongation., Stem Cells 35, 1579-1591, 2017
Human角膜基质细胞干细胞 / Corneal stroma stemCollagenase Type 1: 0.1%DMEMSidney, L., Branch, M., Dua, H. and Hopkinson, A.: Effect of Culture Medium on Propagation and Phenotype of Corneal Stroma-Derived Stem Cells., Cytotherapy 17, 1706-22, 2015
Human角膜基质细胞干细胞 / Corneal stromal stemNeutral Protease: 1.2 u/ml;
Collagenase: 0.1%
DMEMDu, Y., Roh, D., Funderburgh, M., Mann, M., Marra, K., Rubin, J., Li, X. and Funderburgh, J.: Adipose-Derived Stem Cells Differentiate to Keratocytes In Vitro., Mol Vis 16, 2680, 2010
Human, 18-68 yr角膜内皮 / Corneal endothelialCollagenase: 0.2%;
Neutral Protease: 1.0%
DMEM/F12Li Wei, Sabater Alfonso L, Chen Ying-Ting, Hayashida Yasutaka, Chen Szu-Yu, He Hua, Tseng Scheffer C G: A novel method of isolation, preservation, and expansion of human corneal endothelial cells, Inv Ophthal Visual Sci 48, 614-20, 2007
Human角膜缘上皮片 / Corneal limbal epithelial sheetNeutral Protease: 5%see referenceEspana EM, Romano AC, Kawakita T, Di Pascuale M, Smiddy R, and Tseng SC: Novel enzymatic isolation of an entire viable human limbal epithelial sheet, Inv Ophthal Visual Sci 44(10), 4275, 2003
Human, 5-65 years视网膜色素上皮 (RPE) / 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
Macaques and baboon视网膜 / RetinalPapain: 20-40 u/ml;
Deoxyribonuclease I: 400 u/ml
Ames' solutionHan ,Y., Jacoby, R. and Wu, S.: Morphological and Electrophysiological Properties of Dissociated Primate Retinal Cells., Brain Res 875, 175, 2000
Monkey, cynomolgus, young adult结膜淋巴细胞 / Conjunctival lymphocytesCollagenase Type 1: 0.02%RPMI 1640Whittum-Hudson JA, Taylor HR: Antichlamydial specificity of conjunctival lymphocytes during experimental ocular infection, Infect Immun 57, 2977, 1989
小鼠Mouse, 2-4 week米勒细胞 / MullerPDS kit: per instructionsDMEMLiu, X., Tang, L. and Liu Y.: Mouse Muller Cell Isolation and Culture., Bio-protocol 7, , 2017
小鼠Mouse视网膜 / RetinalPapain: 20 u/ml;
Deoxyribonuclease I: 0.005%
PBSOrtin-Martinez, A., Tsai, E., Nickerson, P., Bergeret, M., Lu, Y., Smiley, S., Comanita, L. and Wallace, V.: A Reinterpretation of Cell Transplantation: GFP Transfer From Donor to Host Photoreceptors., Stem Cells 35, 932-939, 2017
小鼠Mouse视网膜 / RetinalPapain: 20 u/ml;
Deoxyribonuclease I: 0.005%
HBSSWohl, S., Jorstad, N., Levine, E. and Reh, T.: Muller Glial microRNAs are Required for the Maintenance of Glial Homeostasis and Retinal Architecture., Nat Commun 8, 1603, 2017
小鼠Mouse, C57BL/6视网膜星形胶质细胞 / Retinal astrocytesPapain: 16.5 u/ml;
Deoxyribonuclease I: 0.004%
HBSSBlandford, S. and Baldridge, W.: The Effect of Glutamate Receptor Agonists on Mouse Retinal Astrocyte [Ca(2+)]i., Biomed Res Int 2016, 8178162, 2016
小鼠Mouse, adult视网膜感光器 / Retinal photoreceptorPDS kit: with modificationsEBSSFeodorova, Y., Koch, M., Bultman, S., Michalakis, S. and Solovei, I.: Quick and Reliable Method for Retina Dissociation and Separation of Rod Photoreceptor Perikarya from Adult Mice., MethodsX 2, 39, 2015
小鼠Mouse, 4-8 day光感受器细胞 / PhotoreceptorsPDS kit: per instructionsNeurobasalBalmer, J., Zulliger, R., Roberti, S. and Enzmann, V.: Retinal Cell Death Caused by Sodium Iodate Involves Multiple Caspase-Dependent and Caspase-Independent Cell-Death Pathways., Int J Mol Sci 16, 15086-103, 2015
小鼠Mouse, embryonic and P0视网膜 / RetinalPapain: 8 u/ml;
Deoxyribonuclease I: 124 u/ml
DPBSDvoriantchikova, G., Perea-Martinez, I., Pappas, S., Barry, A., Danek, D., Dvoriantchikova, X., Pelaez, D. and Ivanov, D.: Molecular Characterization of Notch1 Positive Progenitor Cells in the Developing Retina., PLoS ONE 10, e0131054, 2015
小鼠Mouse玻璃体细胞 / Vitreous cellsCollagenase: 0.1%;
Hyaluronidase: 0.03%
DMEMIqbal, N., Xu, L., Devitt, C. and Skapek, S.: Isolation and Characterization of Mammalian Cells Expressing the Arf Promoter During Eye Development., Biotechniques 56, 239-49, 2014
小鼠Mouse, 6 week视网膜 / RetinalPDS kit: per instructionsDMEMSingh, M., Issa, P, Butler, R., Martin, C., Lipinski, D., Sekaran, S., Barnard, A. and Maclaren, R.: Reversal of End-Stage Retinal Degeneration and Restoration of Visual Bunction by Photoreceptor Transplantation., Proc Natl Acad Sci U S A , , 2013
小鼠Mouse视网膜神经节 / Retinal ganglionCLSPA: 240 u/ml;
Hyaluronidase: 0.2%
AmesSchmidt, T. and Kofuji, P.: An Isolated Retinal Preparation to Record Light Response from Genetically Labeled Retinal Ganglion Cells., J Vis Exp 47, 2367, 2011
小鼠Mouse, adult纤毛上皮 / Cilary epithelialPDS kit: per instructionsEBSSGualdoni, S., Baron, M., Lakowski, J., Decembrini, S., Pearson, R., Ali, R. and Sowden, J.: Isolation and Culture of Adult Ciliary Epithelial Cells, Previously Identified as Retinal Stem Cells, and Retinal Progenitor Cells., Curr Protoc Stem Cell Biol Chapter 1, Unit 1H.4, 2011
小鼠Mouse, 4-5 week透镜纤维 / Lens FiberCollagenase Type 4: 0.125%;
Protease: 0.5%
PBSEbihara, L., Tong, J., Vertel, B., White, T. and Chen, T.: Properties of Connexin 46 Hemichannels in Dissociated Lens Fiber Cells., Inv Ophthal Visual Sci 52, 882-9, 2011
小鼠Mouse, postnatal day 1视网膜祖细胞 / Retinal progenitorCollagenase: 0.1%HBSSJiang, C. Klassen, H., Zhang, X. and Young, M.: Laser Injury Promotes Migration and Integration of Retinal Progenitor Cells into Host Retina., Mol Vis 16, 983, 2010
小鼠Mouse, 4-8 week角膜上皮 / Corneal epithelialNeutral Protease: 1.5%;
Trypsin: 0.25%
DMEM/F12Kobayashi, T., Yoshioka, R., Shiraishi, A. and Ohashi, Y.: New Technique for Culturing Corneal Epithelial Cells of Normal Mice., Mol Vis 15, 1589-93, 2009
小鼠Mouse视网膜 / RetinalCollagenase: 0.1%;
Deoxyribonuclease I: 0.001%
RPMIAmadi-Obi, A., Yu, C., Liu, X., Mahdi, R., Clarke, G., Nussenblatt, R, Gery, I., Lee, Y. and Egwuagu, C.: TH17 Cells Contribute to Uveitis and Scleritis and are Expanded by IL-2 and Inhibited by IL-27/STAT1., Nat Med 13, 711, 2007
小鼠mouse, adult视网膜 / RetinalPapain: 50 u/mlDMEM/F-12Jadhav AP, Cho SH and Cepko CL: Notch activity permits retinal cells to progress through multiple progenitor states and acquire a stem cell property, Proc Natl Acad Sci U S A 103, 18998, 2006
小鼠Mouse, adult基质细胞 / StromalTrypsin: 0.05%;
Collagenase: 78 u/ml;
Hyaluronidase: 38 u/ml
DMEM/F-12Yoshida, S., Shimmura, S., Nagoshi, N., Fukuda, K., Matsuzaki, Y., Okano, H. and Tsubota, K.: Isolation of Multipotent Neural Crest-Derived Stem Cells from the Adult Mouse Cornea., Stem Cells 24, 2714-22, 2006
小鼠Mouse, 2-5 month小鼠视网膜和双极 / Mouse retinal and bipolarPapain: 20 u/ml;
Deoxyribonuclease I: 200 u/ml
HBSSMaxeiner Stephan, Dedek Karin, Janssen-Bienhold Ulrike, Ammermuller Josef, Brune Hendrik, Kirsch Taryn, Pieper Mario, Degen Joachim, Kruger Olaf, Willecke Klaus, Weiler Reto: Deletion of connexin45 in mouse retinal neurons disrupts the rod/cone signaling pathway between AII amacrine and ON cone bipolar cells and leads to impaired visual transmission, J Neurosci 25, 566-76, 2005
小鼠Mouse, 4 week视网膜内皮细胞 / Retinal endothelial cellsCollagenase Type 1: 0.1%DMEMSu X, Sorenson CM, and Sheibani N.: Isolation and characterization of murine retinal endothelial cells, Mol Vis 9, 171, 2003


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