外周血epcs的体外培养与aspirin对epcs迁移功能影响的.研究

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1、天津医科大学硕士学位论文2．TheeffectofaspirinonmigratoryactivityofEPCs：Atthebasementofthefirststepofstudy，cellswhichculturein7daysweredissociated，resuspensed，countedandputsuspensioninuppercompartmentsofTranswellchamber．AndthenputEndoCultLiquidMediumKitwithFBScontainsaspirinofdifferentconcentrationinl

2、owercompartmentsofTranswellchamber．PuttotalTranswellchamberintocellincubator．After24ll，takeTranswellchamberoutandcountmigratorycellsonthelowsideofmembrane．WeobservetheeffectofaspirinOnmigratoryactivityofEPCsindifferentconcentrationaspirinandthesameconcentrationaspirinindifferenttimes．Res

3、ults：1-2x106MNCswereisolatedinonemlperipheralbloodbyFicolldensitygradientcentrifugationwhenweadjustcentrifugerateto1200rpm．TrypanbluestainingmeasuredtheviabilitiesofMNCswereabout95+3％．Inthemorningofculture,themostofcellssh印edroundoroval，mixedwithsmallamountsoffusiformcells．ApartofroundMN

4、CsattacheddishesbecomefusiformEPCsinthefiRhday．Aboutintheseventhday,cellcoloniescontainsseveralcellsemerged．Cellsbecomelong-spindleforminthetenthday．Afterfourteendays，cellsbeginsenesce，disintegration．ThedetectionofendothelialprogenitorcelllinespecificmarkerVEGFR一2，CD133andvWFarestrongpos

5、itiveinseventhday．nefunctionofEPCsforac·LDL-uptakingandUEA—lbindingwerebothstrongpositiveinseventhday．Wechoseseventhday’cells，usedTranswellchambertoobservetheeffectofaspirinonmigratoryactivityofEPCs，countmigratorycellsunder200xmicroscope．Statisticalanalysisconfirmedthataspirinobviouslyde

6、creasemigratorycellpopulationofEPCsandthesemigratorycellpopulationsobviouslydecreasewhenaspirin’Sconcentrationincrease．Conclusions：1．At25-37"C．MNCscallbebetterisolatedfromperipheralbloodwhenuse1200rpmbyFicolldensitygradientcentrifugation．ItCallsatisfythecultureofEPCs．2．Theidentificationo

7、fphenotypeandmensurationofactivityforEPCsexhibittheperiodofprosperityintheseventhdaytotenthdayaftercultureinvitro．3．IntheTranswellchamberculture，lmmol／LaspirinCaninhibitmigratoryactivityofEPCs，thisactionobviouslydependsonaspirin’concentrationandactiontime．Keywords：Periphe