Feasibility Study On Differentiation Of TM Pluripotent Stem Cells Into Corneal Endothelial Cells

Aim:To isolate,identify and characterize multipotent stem cells from human trabecular meshwork(TM),exploring it’s feasiblity of differentiation into corneal endothelial cells.Method:Human trabecular meshwork multipotent stem cells were isolated from the trabecular meshwork by collagenase A,and cultured on coated or three–dimensional Matrigel in MESCM containing leukemia inhibitory factor,basic fibroblast growth factor,and 5% fetal bovine serum,identified by specific cell markers of trabecular meshwork using RT-Real-time PCR and Immunostaining.Their multi-potency of neural crest progenitors was characterized by tri-lineage differentiation using their respective induction media and characterization kits commercially available.Immunostaining was used to detect the expression of human corneal endothelial cells(HCEC)markers in induced polygonal cells.Results: The cells isolated from human trabecular meshwork expressed specific TM markers such as AQP1,MGP and CHI3L1,at m RNA and protein levels.On Coated Matrigel plastic could be expanded to yield spindle cells and had a growth potential for up to 4 passages with 13 cell doublings.Such cells expressed high embryonic stem cell markers such as Oct4,Sox2 and Nanog,and neural crest cell markers,such as p75 NTR,FOXD3 and Sox9,had the potential to differentiate into a number of terminal cells,such as adipocyte-like,chondrocyte-like and corneal endothelial-like cells.Conclusion:Trabecular meshwork cells are specific niche cells located in Schwalbe’s Ring region,which can be isolated by clonal cultures.Such cells are multipotent stem cells with the ability to differentiate into adipocyte-like,chondrocyte-like and corneal endothelial-like cells.