Fig 1: Corneal endothelial cell associated gene expression of differentiated adherent and sphere HFSC. Analysis of the relatively normalized gene expression of ATPase, ZO-1, COL8 and PITX2.A) Adherent HFSC of passage 1 were seeded on a DLT imprinted collagen in a density of 3000 cells/ mm2 and cultivated for seven days. Gene expression data were normalized using the housekeeping genes GAPDH and B2M. Error bars represent three replicates. Relative gene upregulation was assessed in comparison to Jurkat cells as control cultured on tissue culture plates in a seeding density of 1000 c/cm2. B) Sphere outgrowth HFSC were seeded on a collagen membrane with DLT and cultivated for 21 days. Relative gene upregulation was assessed in comparison to Jurkat cells as control cultured on tissue culture plates in a seeding density of 1000 c/cm2. Significance in comparison to control was analyzed for A) and B) by one way anova testing. * indicates p < 0.05, ** indicates p < 0.01 and *** indicates p < 0.001. C) Immunofluorescence analysis was performed to measure marker expression of PITX, Na/K-ATPase, Col-8 and ZO-1. Sphere grown cells were cultivated on DLT collagen for 14 days. For antibody staining single cells were scraped off the collagen I scaffold. Nuclei were stained with DAPI. Scale bars show 50 µm