Fig. 3: Generation and characterization of CIITA knockout monkey iPSCs and RPE cells.
From: Graft survival of major histocompatibility complex deficient stem cell-derived retinal cells
a Experimental design for transplantation of CIITA+/+ and CIITA-/- miPSC-RPE into 4 healthy monkeys (HM-22 received CIITA+/+ miPSC-RPE; HM-23, HM-24, HM-25 received CIITA-/- miPSC-RPE). Both miPSC-RPE lines were generated from 1121A1 monkey iPSC line (MHC homozygous). Samples from two other healthy monkeys (S2-4 and K177) who received transplantation of wild-type miPSC-RPE cells (derived from 1121A1 and 46a iPSC lines, respectively) in our previous study27 were also used in this study. b The schema of the CIITA knockout production. c CIITA-/- miPSC-RPE cells (right) show polygonal and hexagonal morphologies as well as the control CIITA+/+ miPSC-RPE cells (left). Scale bars: 500 μm (upper) and 100 μm (lower). d Western blots for CIITA expression in CIITA-/- miPSC-RPE with or without rIFN-γ pretreatment: lane 1 and 2, CIITA-/- miPSC-RPE cells; lane 3 and 4, wild type control (CIITA+/+); lane 5 and 6, positive control (PBMC from a monkey). Full-length images of the western blots are in Supplementary Fig. 6. e Expressions of MHC class I and II on the cell surfaces of CIITA-/- miPSC-RPE with or without rIFN-γ stimulation were analyzed by flowcytometry. X-axis: the FITC-intensity of the cells stained with FITC-conjugated anti-human HLA-class I or anti-human HLA-class II. Y-axis: frequency.
