Fig. 1

Comparison of the DPT, viability, and morphology of LSC grown with XSHEM or CnT07. a Evaluation of the viability by trypan blue exclusion assay. Cell viability was higher with XSHEM medium after cell detachment. b DPT did not show differences between both media. c Morphology of a starting culture of LSC cultured in XSHEM, corresponding to one clone in expansion. d Morphology of a starting culture of LSC cultured in CnT07. No clones could be observed. e Morphology of LSC cultured in XSHEM. Cells were small and polygonal. f Morphology of LSC cultured in CnT07. Cells had a cobblestone morphology, but differentiated cells could be observed (arrows). g XSHEM cultures showed stratifications as clones grew. h CnT07 cultures only grew forming monolayers and did not form any stratification. Results are presented as mean ± SE from three independent experiments. Statistical analysis was performed using two-tailed Student’s t tests (***p < 0.001). DPT, doubling population time; LSC, limbal stem cells; XSHEM, xenofree supplemented hormonal medium. Bar = 100 μm for c to g. Bar = 50 μm for h