组织工程材料捕获成体干细胞的影响因素研究

2009643ChinMedBiotechnol,June2009,Vol.4,No.3171DOI:10.3969/cmba.j.issn.1673-713X.2009.03.003··BALB/c23714HE-1OP-1G-CSF369121512CD34-1Sca-112OP-1G-CSFMTTHEOP-191215G-CSF9P0.05OP-1CD34Sca-113.07%±0.19%3.98%±0.15%17.02%±0.12%11.62%±0.44%3.03%±0.11%2.91%±0.14%P0.05G-CSFCD34Sca-112.79%±0.39%4.52%±0.35%10.21%±0.15%P0.05MTTOP-1G-CSFP0.05OP-1(3):171-176[1][2]muscle-derivedstemcellsMDSC[3-5][6][7][8]CD34CD45-1stemcellantigen-1Sca-1glialfibrillaryacidicproteinGFAPalphafetoproteinAFPCD105[9]-1osteogenicprotein-1OP-1-βTGF-β[10][e-mail]2009-01-051722009643ChinMedBiotechnol,June2009,Vol.4,No.3DMEMGibcoHyclonePECD34FITCSca-1Ly6A.2/6E.1BioLegendOP-1071206G-CSF080102MTT2mm×20mm×50mmabsorbablegelatinspongeAGS90%2mm×20mm×50mmpolymeroflacticacidPLA90%FACSCaliburBDKD-TS3CKD-BMKD-HHM315MICROMSynergy™HTBio-TEKALCACCULABSPFBALB/c5~612~15gSCXK20030004SPF0.25mg/mlOP-124~48h1.6mg/5mm2mmBALB/c126237142HEOP-1G-CSFBALB/c30OP-1G-CSF10OP-1OP-10.25mg/mlG-CSF250µg·kg-1·d-1[11]7dG-CSF36912152D-HanksDMEM×1003121×106/mlPBS3PECD34FITCSca-13730minPBS2CD34Sca-13OP-1G-CSF1210%10%DMEM5×104/ml100µl/9633d50ng/mlOP-1OP-150µg/mlG-CSFG-CSF100µl824487296h20µlMTT5g/L374h150µlDMSO2min490nmA3SPSS10.0sx±χ2tP0.05HE37142009643ChinMedBiotechnol,June2009,Vol.4,No.317313691215OP-1G-CSF1OP-191215P0.05G-CSF9HE×200ABC3714DEF3714Figure1ObservationofthecellscapturedbyAGSandPLAimplantedinmiceatdifferenttimes(HE,originalmagnification×200).A,B,CrespectivelyshowingthecellsmorphologycapturedbyAGSondays3,7,and14;D,E,FrespectivelyshowingthecellsmorphologycapturedbyPLAondays3,7,and14.Table1QualityofAGStreatedindifferentwaysafterimplantedinmiceatdifferenttimesgQualityofAGS(g)Groups3Day36Day69Day912Day1215Day15OP-1OP-1group0.11980.11710.11410.10750.0872G-CSFG-CSFgroup0.11870.11780.11040.07730.0725Controlgroup0.12010.11680.11460.10120.0921sx±Table2NumberofthecellscapturedbyAGStreatedindifferentwaysafterimplantedinmiceatdifferenttimes(sx±)×106Numberofthecapturedcells(×106)Groups3Day36Day69Day912Day1215Day15OP-1OP-1group0.88±0.041.86±0.052.91±0.19a4.49±0.19a5.96±0.22aG-CSFG-CSFgroup0.84±0.021.23±0.212.36±0.07a3.16±0.363.81±0.14Controlgroup0.71±0.061.15±0.131.45±0.163.11±0.143.67±0.85aP0.05Note:aComparedwithcontrolgroup,P0.05.1742009643ChinMedBiotechnol,June2009,Vol.4,No.312CD34Sca-1Figure2FlowcytometryshowingtheexpressionofCD34andSca-1onthecellscapturedbyAGStreatedindifferentwaysafterimplantedinmiceonday12P0.0512152OP-1CD34Sca-113.07%±0.19%3.98%±0.15%17.02%±0.12%11.62%±0.44%3.03%±0.11%2.91%±0.14%P0.05G-CSFCD34Sca-112.79%±0.39%4.52%±0.35%10.21%±0.15%CD34Sca-1P0.052MTTOP-1G-CSFP0.053OP-1G-CSFA490A490value0.80.60.40.2024487296hProliferationtime(h)Figure3Effectsofdifferentculturemediumontheproliferativeactivityofthecapturedcells[12][13]G-CSFG-CSF5~7[14]CD34G-CSF9OP-1OP-1OP-1CD34Sca-1OP-12009643ChinMedBiotechnol,June2009,Vol.4,No.3175OP-1[1]BrazeltonTR,RossiFM,KeshetGI,etal.Frommarrowtobrain:expressionofneuronalphenotypesinadultmice.Science,2000,290(5497):1775-1779.[2]AndersonDJ,GageFH,WeissmanIL.Canstemcellscrosslineageboundaries?NatMed,2001,7(4):393-395.[3]CaoB,ZhengB,JankowskiRJ,etal.Musclestemcellsdifferentiateintohaematopoieticlineagesbutretainmyogenicpotential.NatCellBiol,2003,5(7):640-646.[4]ArrieroM,BrodskySV,GealekmanO,etal.Adultskeletalmusclestemcellsdifferentiateintoendotheliallineageandamelioraterenaldysfunctionafteracuteischemia.AmJPhysiolRenalPhysiol,2004,287(4):F621-F627.[5]Qu-PetersenZ,DeasyB,JankowskiR,etal.Identificationofanovelpopulationofmusclestemcellsinmice:potentialformuscleregeneration.JCellBiol,2002,157(5):851-854.[6]McKinney-FreemanSL,JacksonKA,CamargoFD,etal.Muscle-derivedhematopoieticstemcellsarehematopoieticinorigin.ProcNatlAcadSciUSA,2002,99(3):1341-1346.[7]TsuboiK,KawadaH,TohE,etal.Potentialandoriginofthehematopoieticpopulationinhumanskeletalmuscle.LeukRes,2005,29(3):317-324.[8]LiJ,WangS,HanJ,etal.Cellscapturedfromspatiumintermuscularebyporousmaterialexhibitthecharacteristicsofstemcells.HistochemCellBiol,2008,130(4):741-748.[9]AlessandriG,PaganoS,BezA,etal.Isolationandcultureofhumanmuscle-derivedstemcellsabletodifferentiateintomyogenicandneurogeniccelllineages.Lancet,2004,364(9448):1872-1883.[10]WangGD,WangSL,HanJX.Advancesinthestudyofbonemorphogeneticprotein-7.JMedMolBiol,2004,1(6):382-385.(inChinese),,.-7.,2004,1(6):382-385.[11]HartungG,UharekL,ZeisM,etal.Superiorantileukemicactivityofmurineperipheralbloodprogenitorcell(PBPC)graftsmobilizedbyG-CSFandstemcellfactor(SCF)ascomparedtoG-CSFalone.BoneMarrowTransplant,1998,21Suppl3:S16-S20.[12]LuL,PeterSJ,LymanMD,etal.Invitrodegradationofporouspoly(L-1acticacid)foams.Biomaterials,2000,21(15):1595-1605.[13]JacobsenS,FritzHG,DegéeP,etal.Newdevelopmentsontheringopeningpolymerizationofpolylactide.IndCropsProducts,2000,11(3):265-275.[14]WangCB,ChenXH,OuYX,etal.DynamicchangesofCD34~+cellandTlymphocytesubsetfromdifferentpopulationsafterG-CSFtreatment.ActaAcadMediMilitTertiae,2005,27(12):1269-1271.(inChinese),,,.G-CSFCD34~+T.,2005,27(