體外誘導人羊膜干細胞分化為胰島素分泌細胞.pdf

1、遵義醫(yī)學院碩士學位論文體外誘導人羊膜干細胞分化為胰島素分泌細胞姓名：趙玉潔申請學位級別：碩士專業(yè)：生物化學與分子生物學指導教師：陳代雄20100501遵義醫(yī)學院碩士學位論文體外誘導人羊膜干細胞分化為胰島素分泌細胞InducingDifferentiationofHumanAmnionStemCellsintoInsulinSecretingCellsinVitroAbstract：Objectives：Somaticstemcell(S

2、SC)transplantationhasreceivedmoreattentionasanewstrategyoftherayfortypeIdiabetesmellitusHumanamnionderivedstemcells(hADSCs)maybeapotentialcellsourceInthisstudy，weinducedhADSCincludinghumanamnioticmesenchymalcells(hAMCs)a

3、ndhumanamnioticepithelialcells(hAECs)todifferentiateintoinsulinsecretingcells(ISCs)invitrotoexploreapotentialnewtherapyfortypeIdiabetesMethods：@hAECsandhAMCswereisolatedfromamniotictissuewithtrypsinandcollagenase，andthep

4、henotypeoftheisolatedcellswasidentifiedbyflowcytometryandimmunocytochemicalstaining②hAECsandhAMCsweredividedintoinducedgroupandnoninducedgroupInducedgroupwastreatedinserumfreeHGDMEMwith10mMnicotinamideandN2supplementtoin

5、vestigatetheirdifferentiationintoISCsThemediumusedinnoninducedgroupWasonlysupplementedwith10％fetalbovineseiumLGDMEM③Angroupswereculturedinahumidifiedatmosphereof95％airand5％C02at37。CAt7，14and21days，theexpressionofinsulina

6、ndp2microglobulinWasdeterminedbyimmunocytochemicalstaining，whilethecontentofinsulininsupernatantwasdetectedbyradioimmunoassayandtheexpressionofInsulinandPDX一1mRNAwasdetectedbyreversetranscfiptasepolymerasechainreaction(R

7、TPCR)Results：①FreshlyisolatedhAECsshowedlowexpressionforCD44andwerepositiveforCK19，buthAMCsshowedhigherexpressionforCD44andwerepositiveforvimentinBothhAECsandhAlVlCswerenegativeforCD34andCD45②At7，14and21days，thepercentag

8、esofinsulinpositivecellsinhAECsinducedgroupswere7400a：173％，7533115％and7567士058％respectively，andthepercentagesofinsulinpositivecellsinhAMCswere7467士153％。7500100％and7433153％respectively，butinnon—inducedgroupswerenegativeBo