人胚胎視網(wǎng)膜干細胞的分離培養(yǎng)與誘導分化.pdf

1、中南大學碩士學位論文人胚胎視網(wǎng)膜干細胞的分離培養(yǎng)與誘導分化姓名：胡蓉申請學位級別：碩士專業(yè)：眼科學（視網(wǎng)膜?。┲笇Ы處煟焊吡?0050501ABSTRACTPurposes1Toestablishaculturesystemofhumanfetalretinalstemcells(RSCs)invitro2Toinvestigatetheroleofretinoicacid(RAlintheinduceddifferentiationo

2、fRSCsinvitroMethods1Retinalstemcellswereisolated丹omhumanfetalneuralretinaeatthe8th12thweekofgestationandculturedrespectivelyinuncoateddishesordishesprecoatedwithpoly—L—ornithineandlaminin(PO／LM)Morphologicalobservationan

3、dimmunohistochemicalanalysiswereapplied2RSCswereculturedinthreedifferentgroups1％fetalbovineserum(controlgroup)，5％fetalbovineserum(5％FBSgroup)and500nMRAplusI％FBS(RAgroup)toinduceRSCstodifferentiateThepositive—expressingce

4、llsandnegetivecellswereidentifiedandcalculatedbymorphologicalandimmunocytochemicalanalysiswithspecificantibodiesagainstNestin，GFAP13Tubulin，RecoverinandRhodopsinnledifferencesoftheratioofpositive—expressingcellsagainstea

5、chantigenwereanalyzedbetweendifferentgroupsResults1TheisolatedcellsformedneurospheresinuncoateddishesandexpressedtheneuroectodermalmarkerNestinbutfailedinpassageⅥ，llilethecellswerecapableofexpressingNestinaswellasdividin

6、gformultiplegenerationsintheprecoateddishes2DifferentiatedRSCsinthethreegroupsexpressedGFAPB—TubulinandRecoverinRSCsdifferentiatedinl％FBS500nMRAmediummaynotonlyexpressthoseabovemarkers，butalshodopsin，withthehighestpositi