Figure 2.

Dental pulp cells (DPCs) differentiate into keratocytes in vitro. DPCs were cultured in keratocyte differentiation medium as pellets for 2 weeks, as described in Materials and Methods. (A): Results of quantitative real-time polymerase chain reaction comparing expression of keratocyte genes in the DPCs after pellet culture or in primary human keratocytes. The expression level in undifferentiated DPCs was set to 1. (B): Immunoblot for keratan sulfate in proteoglycans isolated from conditioned pellet culture media, before and after keratanase treatment. (C): Immunostaining of sections of DPC pellets after 2 days of culture in growth medium for type I collagen (green), with a no-primary antibody control (C′). (D): Immunostaining of sections of DPC pellets after 2 weeks of culture in keratocyte differentiation medium for type I collagen (green), with a corresponding negative control (D′). (E): Immunostaining for keratocan (green) after 2 days of culture in growth medium, with a corresponding negative control (E′). (F): Immunostaining for keratocan (green) after 2 weeks of culture in keratocyte differentiation medium, with a corresponding negative control (F′). Blue represents nuclei stained with 4′,6-diamidino-2-phenylindole. Scale bars = 20 µm. Error bars represent the 95% confidence interval of data.