piD4 host cell
monkey blastocysts after injection of pESCs were cultured on 8-well IbiTreat -plates (IB-80826; Ibidi GmbH) with in vitro culture media in 5% CO2 incubator. For the first 48 h the embryos were cultured in in vitro culture medium 1 (IVC1). On in vitro culture day 2 (embryo day 7 post fertilization), 50% of the culture medium was replaced with in vitro culture medium 2 (IVC2). From in vitro culture day 3 on, IVC2 medium was used and medium was refreshed on a daily basis.For the 96 h, the single host cell were isolated. Embryos were digested into single cells following treatment with 0.5% Trypsin. Cells were grouped with GFP negative or positive using inverted fluorescence microscope. GFP+ injected cells and GFP- host embryonic cells were collected for gene expression analysis, respectively. To exclude GFP cell marker expression used in PCR may come from recipient embryonic cells, single GFP+ cells were picked up under fluorescence microscopy, respectively. The contaminations of the host embryonic cells were excluded before performing RNA isolation. cDNAs synthesis and amplification were performed with Discover-sc WTA Kit V2 (Vazyme N711, China) according to the instruction manual. The qualities of the cDNAs were evaluated by Bioanalyzer 2100.