Time-Mated Breeding to Acquire Rhesus Macaque Embryos at Carnegie Stage 23
(A) Strategy for rhesus macaque time-mated breeding using estrogen to monitor ovulation. Day 1 of embryo development (fertilization) was estimated to occur 72 hr after the estrogen peak.
(B) Estradiol measurements (pg/mL) in the serum of females (animal identification number is shown, n = 10 animals).
(C) Embryo crown rump length (mm) of Carnegie stage 23 embryos (n = 10 embryos).
(D) Embryo weight (g) of Carnegie stage 23 embryos (n = 10 embryos).
(E) A pair of embryonic testicles (T) isolated from a day 47 embryo, and associated mesonephros (m), indicated by the arrow.
(F) H&E stain of a testis at embryonic day 49 and associated mesonephros (m) with glomeruli. Scale bar, 200 μm.
(G) H&E stain of testis cords at embryonic day 49 (arrow points to cords). Scale bar, 50 μm.
(H) Immunofluorescence of embryonic testis at day 49 showing that cKIT-positive PGCs are depleted in 5mC (n = 1 embryonic testis, n = 2 technical duplicates). Scale bar, 10 μm.
(I) Immunofluorescence of embryonic testis at day 49 showing cKIT/OCT4 double-positive PGCs (n = 1 embryonic testis, n = 2 technical duplicate). Scale bar, 20 μm.