worthington 眼睛組織分離指南

發(fā)布者：艾美捷科技發(fā)布時(shí)間：2022-06-01

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原代細(xì)胞分離中，針對眼睛組織涉及到視網(wǎng)膜內(nèi)皮、腦內(nèi)皮細(xì)胞、視網(wǎng)膜細(xì)胞和角膜上皮、米勒細(xì)胞等等的解離通常會使用酶，如何選擇合適的酶來完成對應(yīng)腦組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對眼睛相關(guān)的組織分類解離分種屬、不同的細(xì)胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻(xiàn)，詳細(xì)匯總指南：

眼睛組織分離指南

本worthington 眼睛組織分離指南描述了標(biāo)準(zhǔn)實(shí)驗(yàn)室程序；為建立細(xì)胞分離方案提供了一種合乎邏輯的實(shí)驗(yàn)方法；并列出了許多組織特異性參考資料，以幫助開發(fā)有效的方法。更多其他組織的解離指南，點(diǎn)擊查看：各組織分離指南。

眼睛
種屬	種屬詳情	細(xì)胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻(xiàn)
牛	Bovine	視網(wǎng)膜內(nèi)皮 / Retinal endothelial	Collagenase Type 4: 0.05%； Deoxyribonuclease I: 0.02%； Pronase: 0.02%	DMEM	Terlizzi, 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
牛	Bovine	周細(xì)胞 / Pericyte	Collagenase: 0.2%	DMEM	Bryan, B. and D'Amore, P.: Pericyte Isolation and USe is Endothelial/Pericyte Coculture Models, Meth Enzymol 443, 315, 2008
牛	Bovine	微血管內(nèi)皮 / Microvascular endothelial	Collagenase/Dispase: 0.1%	MEM	Bowman, P., Betz, A., and Goldstein, G.: Primary Culture of Microvascular Endothelial Cells From Bovine Retina, In Vitro 18 (7), 626, 1982
雞	Chick, embryo, 6 day	視網(wǎng)膜細(xì)胞 / Retinal cells	Trypsin: 0.005%； Deoxyribonuclease I: 0.005%	DMEM/F12	Jacob 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
雞	Chick embryo	角膜上皮 / Corneal epithelia	Collagenase: 0.08%； Trypsin: 0.08%	HBSS	Reenstra 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
雞	Chick embryo (also rat)	視網(wǎng)膜 / Retinal	Trypsin: 0.6%	DMEM	Seigel, G.: The Golden Age of Retinal Cell Culture, Mol Vis 5, 4, 1999
雞	Chick, embryo, 10-14 day	平坦，視網(wǎng)膜 / Flat, retina	Trypsin: 0.1%	Tyrode's solution, CMF	Moyer, 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
魚	Fish, zebrafish	視網(wǎng)膜 / Retinal	Papain: 10 u/ml	L-15	Diekmann, H., Kalbhen, P. and Fischer, D.: Characterization of Optic Nerve Regeneration using Transgenic Zebrafish., Front Cell Neurosci 9, 118, 2015
魚	Goldfish	視網(wǎng)膜雙極 / Retinal Bipolar	Papain: 30 u/ml	HEPES	Graffe, 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
人	Human	角膜基質(zhì)細(xì)胞角質(zhì)細(xì)胞 / Corneal stromal keratocytes	Collagenase Type 1: 0.1%	DMEM/F-12	Yusoff,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
人	Human	米勒細(xì)胞 / Muller glia	PDS kit: per instructions	DMEM	Xu, 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
人	Human	角膜基質(zhì)細(xì)胞干細(xì)胞 / Corneal stroma stem	Collagenase Type 1: 0.1%	DMEM	Sidney, 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
人	Human	角膜基質(zhì)細(xì)胞干細(xì)胞 / Corneal stromal stem	Neutral Protease: 1.2 u/ml； Collagenase: 0.1%	DMEM	Du, 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
人	Human, 18-68 yr	角膜內(nèi)皮 / Corneal endothelial	Collagenase: 0.2%； Neutral Protease: 1.0%	DMEM/F12	Li 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
人	Human	角膜緣上皮片 / Corneal limbal epithelial sheet	Neutral Protease: 5%	see reference	Espana 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
人	Human, 5-65 years	視網(wǎng)膜色素上皮 (RPE) / Retinal pigment epithelial (RPE)	Trypsin: 0.25%	HBSS	Von 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
猴	Macaques and baboon	視網(wǎng)膜 / Retinal	Papain: 20-40 u/ml； Deoxyribonuclease I: 400 u/ml	Ames' solution	Han ,Y., Jacoby, R. and Wu, S.: Morphological and Electrophysiological Properties of Dissociated Primate Retinal Cells., Brain Res 875, 175, 2000
猴	Monkey, cynomolgus, young adult	結(jié)膜淋巴細(xì)胞 / Conjunctival lymphocytes	Collagenase Type 1: 0.02%	RPMI 1640	Whittum-Hudson JA, Taylor HR: Antichlamydial specificity of conjunctival lymphocytes during experimental ocular infection, Infect Immun 57, 2977, 1989
小鼠	Mouse, 2-4 week	米勒細(xì)胞 / Muller	PDS kit: per instructions	DMEM	Liu, X., Tang, L. and Liu Y.: Mouse Muller Cell Isolation and Culture., Bio-protocol 7, , 2017
小鼠	Mouse	視網(wǎng)膜 / Retinal	Papain: 20 u/ml； Deoxyribonuclease I: 0.005%	PBS	Ortin-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	視網(wǎng)膜 / Retinal	Papain: 20 u/ml； Deoxyribonuclease I: 0.005%	HBSS	Wohl, 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	視網(wǎng)膜星形膠質(zhì)細(xì)胞 / Retinal astrocytes	Papain: 16.5 u/ml； Deoxyribonuclease I: 0.004%	HBSS	Blandford, 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	視網(wǎng)膜感光器 / Retinal photoreceptor	PDS kit: with modifications	EBSS	Feodorova, 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	光感受器細(xì)胞 / Photoreceptors	PDS kit: per instructions	Neurobasal	Balmer, 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	視網(wǎng)膜 / Retinal	Papain: 8 u/ml； Deoxyribonuclease I: 124 u/ml	DPBS	Dvoriantchikova, 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	玻璃體細(xì)胞 / Vitreous cells	Collagenase: 0.1%； Hyaluronidase: 0.03%	DMEM	Iqbal, 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	視網(wǎng)膜 / Retinal	PDS kit: per instructions	DMEM	Singh, 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	視網(wǎng)膜神經(jīng)節(jié) / Retinal ganglion	CLSPA: 240 u/ml； Hyaluronidase: 0.2%	Ames	Schmidt, 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	纖毛上皮 / Cilary epithelial	PDS kit: per instructions	EBSS	Gualdoni, 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	透鏡纖維 / Lens Fiber	Collagenase Type 4: 0.125%； Protease: 0.5%	PBS	Ebihara, 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	視網(wǎng)膜祖細(xì)胞 / Retinal progenitor	Collagenase: 0.1%	HBSS	Jiang, 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 epithelial	Neutral Protease: 1.5%； Trypsin: 0.25%	DMEM/F12	Kobayashi, 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	視網(wǎng)膜 / Retinal	Collagenase: 0.1%； Deoxyribonuclease I: 0.001%	RPMI	Amadi-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	視網(wǎng)膜 / Retinal	Papain: 50 u/ml	DMEM/F-12	Jadhav 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	基質(zhì)細(xì)胞 / Stromal	Trypsin: 0.05%； Collagenase: 78 u/ml； Hyaluronidase: 38 u/ml	DMEM/F-12	Yoshida, 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	小鼠視網(wǎng)膜和雙極 / Mouse retinal and bipolar	Papain: 20 u/ml； Deoxyribonuclease I: 200 u/ml	HBSS	Maxeiner 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	視網(wǎng)膜內(nèi)皮細(xì)胞 / Retinal endothelial cells	Collagenase Type 1: 0.1%	DMEM	Su X, Sorenson CM, and Sheibani N.: Isolation and characterization of murine retinal endothelial cells, Mol Vis 9, 171, 2003