Journal: STAR Protocols
Article Title: An improved protocol for generation and characterization of human-induced pluripotent stem cell-derived retinal pigment epithelium cells
doi: 10.1016/j.xpro.2022.101803
Figure Lengend Snippet: Characterization of RPE (A–E) Immunostaining of iPSC derived RPE cultures for the marker expression during stages of retinal differentiation. Images showing the expression of RPE commitment marker MITF (A), melanocyte protein- PMEL17 (B), RPE maturation markers TYRP1, RPE65 and TYR (C-E)and tight junction marker ZO-1 (D and E). (F) Flow cytometry analysis of iPSC derived RPE cells for positive (MITF, RPE65, TYRP1) and negative (OCT4) markers. (G and H) Gene expression analysis using qPCR for selected markers represented as fold change compared to iPSC. All the positive markers (G) have higher expression and the negative markers (H) show a negligible expression in iPSC derived RPE. (I) ELISA based quantification of secreted PEDF from the in vivo culture supernatant from apical, basal and 2D RPE cultures. Images and graphs are representative of minimum three independent experiments. Scale bars 100 μm.
Article Snippet: MiTF Recombinant Rabbit Monoclonal Antibody (JF100-01) (1:150) , Novus Biologicals , MA5-32554.
Techniques: Immunostaining, Derivative Assay, Marker, Expressing, Flow Cytometry, Gene Expression, Enzyme-linked Immunosorbent Assay, In Vivo