成纤维细胞与完全脱细胞人羊膜的三维黏附

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1、大津医科人学硕十学位论文焦现象。HFF种植在TAHAM上8～14h后细胞移动速度可达12t．tm／h，当细胞在原位形成细胞团块时移动速度接近0I_tm／h。在TAHAM上细胞的移动轨迹接近直线，而在普通培养皿上的细胞移动为无序随意运动，移动速度为16~481．tm／h。结论：TAHAM保留了羊膜的原有主要成分和力学特征，HFF可在TAHAM上形成3D黏附，其细胞形态及细胞移动方式符合3D黏附特征。关键词：羊膜细胞黏附整合素类成纤维细胞3D黏附ll天津医科人学硕+学位论文Abstractobjective：Paxillinanda5integrincoloc

2、alizedtocell—matrixattachmentsparalleltofibronectin-containingextracellularfibers，Thesemoleculestructureswereproposedfortheterm“3D—matrixadhesions．”Thesestructuresincelladhesion，migration，signaling,andcytoskeletalfunctionisdifferentfromstudiesonplanar2Dtissueculturesubstrates．Wepr

3、esentherethehumanforskinfibroblasts(HFF)formed3D-matrixadhesioncomplexesonthethoroughlyacellularhumanamnioticmatrix(TAHAM)．Methods：TAHAMwereproducedbysuspendingdigestionwithtrypsin．TheHFFwereseededon6wellplate，matrigelandTAHAMindividually．11helightmicroscope，scanningelectronicmicr

4、oscope，immunohistochemistryandimmunofluorescencewereusedtoobservethemicro-structuresanddetectionthetypel，III，IV,VIcollagen，laminin，fibronectin，TGF-pl’TGF—p2，FGFoftheTAHAM．Deoxyribonucleicacid(DNA)contentoffreshHAMandTAHAMWritsquantifiedusingHoechst33258dye．Phasecontrastmicroscopew

5、asengagedtoobservethemorphologyofHFEThetimelapseCCDandthetraceanalysissoftwarewereemployedtoprescribethecellmigration．The3Dadhesionfociwereindentifiedbythelaserconfocalmicroscope．ThestrainoftheTAHAMWastestedbytheuniversalmechanicaltestinginstrument．Results：ThefibersoftheTAHAMwerei

6、ntact，typeI，III，IV,VIcollagen，laminin，fibronectinwerepositive，TGF—pl，TGF-p2，FGFwerenegative．HFFwerebipolarextended，spindleshaped，formmultilayercellclustersnetworksandinvadededintothematrix．Hoechst33258dyeconfirmedonTAHAMsurfacewithoutcellsandresidualdebris．TAHAMwascomparedwithfres

7、hamnioticmembraneinthetransitionalstress，thetransitionalstrain，failurestress，failurestrain，thedifferenceWasnotstatisticallysignificant(Pvalues>0．05)(transitionalstress：t=-1．83，P=0．08；transitionalstrain：t=-1．07，P=O．30；failurestress：t=1．57，P=0．13；failurestrain：t=一1．81，P=O．08)．HFFsee

8、dedontheTAHAMfor24h，thecellsexten