提高大鼠脂肪间充质干细胞转化为胰岛样细胞能力的实验研究-肝胆血管外科专业论文.docx

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提高大鼠脂肪间充质干细胞转化为胰岛样细胞能力的实验研究-肝胆血管外科专业论文.docx

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提高大鼠脂肪间充质干细胞转化为胰岛样细胞能力的实验研究-肝胆血管外科专业论文

AbstractObjective:(1)Theseparation,extractionandcultureofratadiposemesenchymalstemcells.(2)Tostudytheeffectsofinjuriedpancreaticextractandcytokinesontheindudceddifferentiationofratadiposemesenchymalstemcells(AMSCs)toislet-likecells.Methods:(1)Theadiposetissueswereseparatedfromratbythesurgicaloperation underasepticcondition.ThecollagenaseandtrypsindigestionwereusedtoextracttheAMSCsfromtissuesandcells’purification.(2)Itconsistedoffourgroups:controlgroup,celldruginducedgroup,mixedinducedgroupandinjuriedpancreaticextractgroup.Differentcytokinesandinjuriedpancreaticextractwereaddedtodifferentgroups.Thecellwasinducedwithdifferentmethodsfor11days.Cells’morphology changeswereobservedandrecordedwithinvertedmicroscope.Insulin-positivecells wereidentifiedbydithizone(DTZ)stainingandtheamountofinsulinofeachgroup afterbeingstimulatedinthehighglucoseenvironmentweremeasuredthrough enzyme-linkedimmunosorbentassay.Therelatedgeneexpressionwasdetectedwith real-timequantitativepolymerasechainreaction(RT-PCR)andimmunofluorescencestaining.Results:(1)TheratAMSCsattachedtotheplate.Theprimarycellswerenearly round.Thespindlecelltypeorcell clustersappeared after36-48h.Cellclusters grew aroundandthegapamongcellsdecreasedgraduallyinthenext7-8days,finally,thecellsarrangedlikewhirlpool.Atabout10days,thecellgrowthdensityreached 70%-80%ofthebottomofbottle.Thecells’purificationbecamehigherthanbeforeaftertrypsindigestion.(2)Thecellsofinducedgroupsgatheredintoclustersandtheir shapesbecameround.DTZstainingpositivecellsappeared.Insulinwassecretedby thecellsunderhighglucoseenvironment,theinsulinlevelandtherelatedgenes expressionofmixedinducedgroupwerehigherthanthoseofotherinducedgroups. Therewerenochangesontheindexesmentionedabove.Conclusion:(1)TheAMSCscanbecollectedthroughcollagenasedigestionand trypsindigestionwasusedforcells’purification.(2)TheinjuriedpancreaticextractcanpromoteAMSCstodifferentiateintoislet-likecellsandthetransformationability ofAMSCscan beimprovedbyit.Keywords:Adiposemesenchymal stem