p21調(diào)控兒童腦腫瘤干細胞的實驗研究.pdf

1、中南大學(xué)碩士學(xué)位論文p21調(diào)控兒童腦腫瘤干細胞的實驗研究姓名：黃永凱申請學(xué)位級別：碩士專業(yè)：神經(jīng)外科指導(dǎo)教師：方加勝20060501碩士學(xué)位論文英文摘要Experimentalstudyonp21regulatingchildrenbraintumorstemcellsABSTRACT0BJECTIvEBraintumorsageamongthemostcommonformsofcancerinchildrenandaccountfor

2、mostcancerrelateddeathsinthisagegroup，Theaimsoftheljr亂partofthepapereretoestablisharemedialsystemtoisolateandpropagatebraintumorstemcells(BTSCs)withinchildrengliomasofdifferenttypesandmedulloblastomas，tostudytheirselfren

3、ewproliferationandmonoclonalformationcharacteristicstodeterminethespecificmakerespeciallyspecificcellsurfacemarkerofBTSCsOneofthetwomainpurposesofthesecondpartofthisinvestigationistolocatep21inthesingleBTSCs，tOanalyzethe

4、differenceofp21inBTSCsanddifferentiatedtumorcells療omBTSCsMETHODSInthisStUdysamplesfrom12gliomasofdifferenttypesand8medulloblastomaswereobtainedfromchildrenpatientsundergoingmicrosur#calllnflorresectionsTumorlnasseswereae

5、utdydissociatedandtrituratedintosinglecellswith025％panereatin，whichwerethenseededintoserumfreeDMEM／F12mediumcontainingB27(1：50)basicfibroblastgrowthfactor(bFGF20n洲，epidermalgrowthfactor(EGF20lag／L)AidertheprimaryBTSsorne

6、urospheresgeneratedtheyweremechanicallydissociatedwith025％panereafinThelimitingdilutionassaywasalsoperformedtoobservetheBTSmonoclonalformationsTheexpressionsofnormalNSCantigenssuchasCDl33andNestinBTSsorneurosphereswereas

7、sayedbyimmunofluoreseenceTheninthesecondpartdifferentiationofBTSCsWaSinducedinmitogenfreeDMEM／F12mediumsupplementedwith10％fetalbovinesenlmTheexpressionsofp21inBTSCsanddifferentiatedtumorcellofBTSCswereassayedbyimmunoeyto

8、chemistryandWesternBlotRESULTSAremedialsystemtoisolateandpropagatewithO25％pancreatiniseasytoobtainBTSCsofchildrenbraintumorsspecia／lyfromlowergradelesionTheexpressionsofCDl33andNestinwereassayedbyimmunofluorescenceincMld