Immunostaining analysis of the expression of progenitor- and proliferation-related markers.

<p>After four days of culture in proliferation conditions, the cells were fixed and immunostained with antibodies against nestin (A-D) and ki-67 (E-H). The ratio of the nestin-positive is similar between different groups, while the percentages of ki-67-positive cells were high in the RPC cultures in BMSC CM, Lens CM and NSC CM, when compared with the cultures in SM condition (I). The percentage of positive cells was determined by dividing the number of immunopositive cells by the number of nuclei stained with DAPI. Five hundred to one thousand cells for each RPC subgroup and each culture were counted in random fields. Scale bars: 50 µm.