Induce Human Umbilical Cord Mesenchymal Stem Cells And Vascular Endothelial Cells Into Corneal Endothelial Like Cells In Vitro

Objective (1) To improve the success rate of hUC-MCs separation in vitro(2) To explore the possibility of hUC-MCs as engineering corneal tissueseed cells(3)To improve the VECs cryopreserving method(4) To explore the possibility of VECs as engineering corneal tissue seedcellsMethods (1) Separate the human umbilical cord-mesenchymal stem cells in vitrowith three methods: Multi-chip of umbilical cord tissue suspended combine with Ⅳcollagenase digestion method,Simple multi-chip of umbilical cord tissue suspendedmethod, umbilical cord tissue suspended method. After isolation,the cells’ shape andsurface markers is observed and detectde through the use of flow cytometry,andinduced to be adipocaytes and osteoblasts.(2)Cultivate the hUC-MSCs on porcine corneal stroma to build tissueengineering corneal lamellar， observe the corneal stroma whether it istransparent,detect it’s surface marker for NSE and ZO-1which is specific for cornealendothelial cells(3)Cryopreserve the VECs with two kinds of methods,compare theiractivity after4weeks through their survival rates,growth curve and apoptosis rate byflow cytometry(4)Cultivate the VECs on porcine corneal stroma to build tissueengineering corneal lamellar,scaned it’s surface structure by electron microscopyResults (1)The success rate of the Multi-chip suspended umbilical cord slicescombined with type IV collagenase digestion method and Simple multi-chip ofumbilical cord tissue suspended method were higher than umbilical cord tissuesuspended method(2)The corneas with hUC-MSCs are transparenter than withoutit,surface-specific endothelial markers NSE and ZO-1expression were positive on hUC-MSCs(3)The reformed method is better than traditional method in morphology,andthere was no significant difference in thei survival rate,growth curve and the apoptosis ratebetween these two kinds of methods(4) Scanning electron microscopy showed the surface of the corneas withVECs grew wellConclusion (1) In this research,we study sever kinds of methods to isolatehUC-MSCs in vitro,and create a more effcient method(2)The hUC-MSCs can converse into CECs(3)The reformed cryopreserved method is convenient and feasible(4)The VECs may be new seed cells to establish engineering corneal...