Effect Of Three-dimensional Co-culture Model On Rabbit Corneal

Objective:To investigate the effect of three-dimensional co-culture model on the expansion of rabbit limbal stem cells in vitro.Methods:The human p eriodontal ligament stem cells(HPDLSCs)was served as the feeder cells fo r rabbit corneal limbal stem cells in vitro,and according to different carriers divided into three groups,the fibrin glue group were co-cultured with fibrin glue,the HPDLSCs,and rabbit limbal stem cells;the amniotic membrane g roup were co-cultured with amniotic membrane,the HPDLSCs,and rabbit Ii mbal stem cells;the control group were co-cultured with the HPDLSCs.Afte r co-coltured for 5 days,we used inverted microscope,hematoxylin and eosi n staining and scanning electron microscopy to observe the cells morpholog y;then the LSCs positive marker p63 expression were compared by immuno fluorescence staining among three groups.Results:Cells in the fibrin glue g roup and the amniotic membrane group were uniform distributed and serrie d,whereas the control group cells were relatively sparse;Scanning electron microscopy showed that the cells were well adhered to the carrier.Immuno fluorescence staining showed that the positive rates of fibrin glue group,th e amniotic membrane group,t-he control group were(69.93±8.76%),(78.36±8.56%),(58.59±8.31%),there was a significant difference among the three gr oups(F=9.43,P=0.00),the positive rate of fibrin glue group and amniotic me mbrane group was higher than that of control g-roup(P<0.05),but there was no significant difference in the positive rate of p63 among the two groups(P>0.05).Conclusion:The co-culture 3D model is helpful f-or ex vivo expans ion of rabbit corneal Iimbal stem cells.