Figure 3. Characterized porcine RPE cells derived from E60 chimeric fetus by complementation of Bama MITF^{L 247S/L247S} embryos with WT blastomeres.

• A
  Representative light microscopic images show that the in vitro‐cultured RPE cells exhibited a hexagonal shape. D3, D5, and D11, cultured for 3, 5, and 11 days, respectively. Scale bars, 100 μm.
• B–D
  Representative immunofluorescence images showed the expression of RPE‐specific markers, including RPE65 (B), ZO‐1 (C), and Bestrophin (D), respectively, in the cultured RPE cells. Scale bars, 50 μm.
• E
  Semi‐quantitative RT–PCR analysis of RPE functional markers in cultured porcine RPE cells. GAPDH was used to confirm the cDNA quality of all the samples.
• F
  Western blotting identified three RPE markers in porcine RPE cells. GAPDH served as a housekeeping gene control.
• G
  ELISA identified secreted VEGF and PEDF in chimeric and WT porcine RPE cells. Media were collected at 48 h after the cells were seeded onto 12‐well cell culture plates to be used for analyses. The results are representative of three independent experiments and were evaluated using a two‐tailed t‐test. No significant differences were identified. Data are presented as the mean ± standard error mean (SEM). WT, RPE cells cultured from the NW‐8 fetus. CH, RPE cells cultured from chimeric NW‐7 fetus.

Source data are available online for this figure.