Figure 4. Generation and Interspecies ICM Incorporation of Different Types of hiPSCs.
(A) Schematic of the strategy for generating naive, intermediate, and primed hiPSCs.
(B) (Top) Representative bright-field images showing the colony morphologies of naive (2iLD-, 4i-, and NHSM-hiPSCs) and intermediate (FAC-hiPSCs) hiPSCs. Bottom, representative immunofluorescence images of naive and intermediate hiPSCs stained with an anti-OCT4 antibody. Red, OCT4; blue, DAPI. Scale bar, 100 μm.
(C) Schematic of the experimental procedures for producing cattle and pig blastocysts obtained from in vitro fertilization (IVF) and parthenoactivation, respectively. Blastocysts were subsequently used for laser-assisted blastocyst injection of hiPSCs. After hiPSC injection, blastocysts were cultured in vitro for 2 days before fixation and analyzed by immunostaining with an anti-HuNu and an anti-SOX2 antibodies. Criteria to evaluate the survival of human cells, as well as the degree and efficiency of ICM incorporation are shown in the blue box.
(D) Number of hiPSCs that integrated into the cattle (left) and pig (right) ICMs after ten hiPSCs were injected into the blastocyst followed by 2 days of in vitro culture. Red line, the average number of ICM-incorporated hiPSCs. Blue dot, the number of ICM-incorporated hiPSCs in each blastocyst.