Clonal analysis of the in vivo differentiation potential of keratinocytes

摘要: Purpose. This study investigated the in vivo differentiation of conjunctival keratinocytes. Methods. Keratinocytes from fornical region conjungtival epithelium were isolated and plated at low density (5 X 102 per 100-mm dish) Dulbecco's minimum essential medium containing 20% fetal bovine serum presence mitomycin C-treated 3T3 feeder cells. At this density, only single, cells attached after overnight culture. Eight days later, small, well-isolated colonies separated one another by detached as a sheet dish injected subcutaneously into flanks BALB/c athyinic mice through an 18-gauge needle. Within day, small firm nodule appeared site injection. different time points, animals killed, nodules excised for morphologic, histogeometrie, cell kinetic analyses. Results. Each implanted colony derived single gave rise to epithelial cyst lined with reconstituted stratified epithelium. Goblet-like loaded periodic acid-Schiff-positive cytoplasmic granules began appear singularly some cysts day 8 postimplantation observed approximately 85% 14. Conclusions. Because formed clonal populations majority had mixed keratinocyte-goblet phenotype, these results suggest strongly existence bipotent precursor that can give both goblet nongoblet system be used factors influence commitment pluripotent stem divergent pathways differentiation.