人脐血来源内皮祖细胞的分离、培养及鉴定(5)

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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