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人脐血来源内皮祖细胞的分离、培养及鉴定(2)

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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

inducedgroupobviouslywhiletherewasalmostnoCDl33expressed.Inuninduced

group,nosuchcellsurfacemarkersexpressedobviously.Compared、vitlltheoriginal,

FACSshowedthatcell

14markerdaysofCDl33decreased(3.1l±1.05)%to(0.09±0.02)%,P<0.05.After

characteristicculture,Weibel-Paladebodies,which眦theonultmslrucuresofendothelialcells,wereshownthetransmission

electronmicroscopephotomicrographsininducedgroup.The

uninducedgroup,inducedgroupandmatureendotheliallevelofNOinthegroupwere(12.43±cells

4.51)umol/L、(52.074-2.17)umol/Land(83.65q-6.14)umoFL,p<0.05.Thelevelof

NOintheinducedgroupWashigherthanthatoftheuninducedgroup,butlowerthan

thatofthematureendothelialcells.Cellsininducedgroupproliferatedmorequickly

thanthatofuuinducedgroupandthetotalnumberofcellsamoutedto105after2

w∞ksofculture.

Conclusion:

1.Mononuclcarcellscouldbeisolatedfromfreshcordbloodby6%HESanddensity

gradientcentrifugationandthereWel'eendothelialprogenitorcellsincordblood

mononuclearcells.

2.Endothelialprogenitorcells

SOonculturedin10%FBSDMEMsupplementedwitIlVEGF,bFGFandcouldbeinducedintoendothelialcells.

bymorphology,markers

toon3.Endothelialprogenitorcellscouldbeidentifiedcellsurface,uitrastruome

proliferate.ofthecells,functionproduceNOandabilityto

4.Endothelialcellsinducedfromcordbloodderivedendothelialprogenitorcellshad

greatabilitytoproliferateandthetotalnumberofcells

numberumoutedoftissueto10S,whichcouldsatisfythe

cardiovascular

possiblyhadneededofastheseedcellsengineeringandthehadpartfunctionofendothelialthecells,SOitindicatedthattheyfeasibilityofasseedcellsoftissueengineering

cardiovascularreplacement.

Keywords:

cordbloodendothelialprogenitorcellsendothelialcellstissueengineering4

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