FIG. 2.

Clonogenicity and growth curve of RW-GFP cells. (A) When cultivated at a low density in liquid PrEGM culture medium in ultra-low attachment 24-well plates in nonadherent conditions, RW-GFP cells formed more spheres than parental RWPE-1 cells; lower panels represent spheres formed after 15 days of culture. (B) When 50–200 cells per well were plated in six-well plates in K-SFM medium, RW-GFP cells formed more colonies than parental RWPE-1 parental cells; lower panels are pictures of 2-weeks-old colonies after fixation and Giemsa staining. (C) RW-GFP cells grew slower than parental RWPE-1 cells; cell growth curves were obtained after plating 2×10⁴ cells per well in six-well plates, and viable cell numbers were determined daily after trypsinization. (A, B, C) Data represent mean values±standard error of the mean of three independent experiments, P values was determined by Student's test ***P<0.001, **P<0.01, and *P<0.05.