Generation of Cynomolgus Monkey Chimeric Fetuses using Embryonic Stem Cells
NHSMV-Cultured cESCs Could Generate Proper Monkey Chimeric Fetus after They Were Reintroduced into Monkey Morulae
(A) Strategies for two different culture protocols (Pr-1 and Pr-2) of morulae after they received cESCs.
(B) The chimeric embryos developed into hatched embryos and maintained cESC integration. pciD1, 3, and 5 indicated one, three and five days (respectively) after cESCs were injected into morulae.
(C) EGFP- and WPRE-specific PCR analysis of embryonic genomic DNA on day 4 after cESC injection. Positive, positive control; Emr1, 2, 3, three different embryos with obvious GFP signal, respectively; Emr4, one embryo without any GFP signal.
(D–F) EGFP- and WPRE- specific PCR analysis (D and E, respectively), and gender-specific gene (SRY) PCR analysis for genomic DNA of the female monkey fetal tissues (F). Pl, placenta; Br, brain; Ey, eye; Sk, skin; Mu, muscle; BM, bone marrow cells; Bo, bone; Te, testis; Sp, spleen; Ki, kidney; He, heart; In, intestine; Bl, bladder; Li, liver; Pa, pancreas; St, stomach; Lu, lung; Ut, uterus; SIV, SIV-GFP vector; ESC, NHSMV-cultured cESCs; SM, surrogate mother; Fa, the father (sperm donor); Mo, the mother (egg donor); Wa, water, negative control.
(G–K) Genetic analysis based on STR examination demonstrating the presence of the cESC DNA haplotype in both fetal tissues. DXS8043 is located in the X chromosome. ND, no detection.
See also Figure S3 and Figure S4 .