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