人脐血来源内皮祖细胞的分离、培养及鉴定(5)
时间:2025-02-24
时间:2025-02-24
人脐血来源内皮租细胞的分离,培养及鉴定
Isolation、cultureandidentifyofhumancordblood
endothelialprogenitorcells
Abstract
objective"
Toinducecordbloodderivedendothelialprogenitorcells(EPCs)into
endothelialcells,investigatethemethodforisolating,culturingandidentilyingEPCs
ofhumancordbloodandthefeasibilityofthesecellsastheseedcellsoftissue
en西neeringcardiovascularreplacement.
MaterialandMethod:
cordbloodweT|eused.MononuclearFifteenhuman
freshcellsⅥ恤isolatedfromcordbloodby6%HESanddensitygradientcentrifogation。T11isstudywas
dividedintotwogroupsaccordingtOdifferentmedium:inducedgroupanduninduced
group.Ininducedgroup.cells
SOonwereculturedininuninducedI∞AFBSDMEMsupplementedwithwereculturedinVEGEbFGFandandgroup,coilsIO%FBS
DMEM.Attachedcellswereidentifiedbymorphologyobservedunderphasecontrast
microscope,characteristicuRrastructureofcellsdetectedbytransmissionelectron
microscope,markersoncellsurface:vWF,VEGFR一2
andflowandCDl33ofdetectedbyimmunofluorescence
oxide(NO)evaluatedstainingcytometry,functionproducingnitricbyquantificationofNOlevelinculturemediumandabilitytO
proliferateobscrvedunderphaseconffastmicroscope.
Result:
ThepercentageofmononuclearcellsisolatedfromfreshcordbloodWas(3.4±
2.1)×107/mL.Ininduced
beinggroup,themorphologyofattachedcellschangedwhileculturedandinduced,fromsmall—sizedroundcellstOspindle-likecells.formcell
clusters.tOtypical“cobblestone”morphologya/k'r
no2weeks.Inuninducedgroup,7daysoftherewerelesscellclustersandtypical"cobblestone”morphology.AfIcr
culture,immunofluorescencestainingshowedthatvWFandVEGFR-2expressedin3
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