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BACKGROUND:Preterm premature rupture of the membrane is a frequent complication during gestational period. Three-dimensional culture can help damaged tissue restore anatomical integrality,being worth for clinical application. OBJECTIVE:To explore biological characteristics of human amniotic cells three-dimensionally cultured by fibrin scaffold,study the possibility of cell/scaffold composite material for repairing preterm premature rupture of the membrane,and to compare with normal monolayer culture. DESIGN,TIME AND SETTING:An in vitro cell/scaffold study was performed at the Basic Institute of Chongqing University of Medical Science between December 2007 and November 2008. MATERIALS:Amnion tissue derived from uterine-incision delivery women was provided by the Department of Obstetrics and Gynecology,the First Affiliated Hospital,Chongqing University of Medical Sciences;fibrinogen was provided by Sigma,USA. METHODS:Either epithelial cells or mesenchymal cells,being isolated from human amniotic membrane,were cultivated using enzyme digestion and repeated adherence methods. Fibrinogen was conglomerated,and epithelial cells of logarithmic growth were plated onto the surface of fibrin scaffold to simulate a three-dimensional culture in vitro. Before conglomerating fibrinogen,mesenchymal cells of logarithmic growth were mixed with fibrin solution to simulate a three-dimensional culture in vivo. In addition,either epithelial cells or mesenchymal cells were incubated onto 24-well plate for normal monolayer culture. MAIN OUTCOME MEASURES:Cell morphology using inverted microscopy,proliferation of mesenchymal cells,and interaction between mesenchymal cells and scaffold under different culture environments. RESULTS:Epithelial cells were round and smooth on the surface of fibrin scaffold,pseudopodia were stretched out,and microvillus were rich. Mesenchymal cells in fibrin glue were fusiform in shape and stretched out along scaffold. Cells formed net structure at different surfaces. By three-dimensional culture,proliferation of mesenchymal cells was stable but slow compared to monolayer culture. By in vivo three-dimensional culture,fibrin glue gradually shrank and gel thickness gradually decreased. On the 5th day,the gel thickness was only 40% for the initial thickness,and on the 15th day,gel thickness was 10%. CONCLUSION:Both epithelial cells and mesenchymal cells can sterically grow on fibrin scaffold,and the proliferation is stable. Fibrin glue gradually shrinks in cell/fibrin composite scaffold,suggesting that amniotic cells/fibrin composite scaffold can simulate body tissues to repair preterm premature rupture of the membrane.