Fig. 4

mNRPCs can differentiate into mature retinal cells in vitro. a–c Morphology of mNRPCs in the sandwich induction system on the 1st (a), 3rd (b), and 5th (c) day. d–f Eight days after sandwich induction, the cells expressed RPE associated genes, including PAX2 (d), ZO-1 and RPE-65 (e), and PAX6 and MITF (f). g–i Cells cultured in 3D condition will form spheres and can differentiate into rod photoreceptor-like cells. Spheres under bright field after 10 days (g) and 3 weeks (h, i). j–o Three weeks after 3D induction, differentiated cells expressed GLYT-1 (j), RHO (j, k), PAX6 and RCVRN (i), RC2 (m), NEUN and PKC-α (n), SYN1 (o), but did not express MITF (k), OPSIN (m), and GFAP (o). Magnification, × 200 (a, b, d–f), × 400 (c, g, i), × 100 (h, j–o); scale bars, 100 μm (a–h, j–o), 50 μm (g, i)