Figure 4
From: Generation of rat offspring using spermatids produced through in vitro spermatogenesis
Offspring production with microinsemination. (a) A P8 rat testis tissue cultured under a PC chip with rat medium 2 for 47 days. BF stands for brightfield. (b) After dissociation, the testis tissue contained round spermatids showing GFP-positive acrosomes (arrowheads). The rectangular areas are enlarged in the bottom panel. (c) Oocytes fertilized with the round spermatids (ROSI), at 24 h after the procedure. On close observation, two pronuclei were identified (arrows). (d) Four pups were delivered by caesarean section. (e) Body weights of the 4 rats were monitored for 12 weeks, showing normal growth. Open circles and triangles indicate body weight of a female and 3 males of ROSI offspring. Green triangles and purple circles represent average body weight ± SD values of normal 20 male and 20 female SD (Crl:CD) rats, respectively, adapted from Kobayashi et al.40. (f) The next generation was produced by natural mating. A ROSI-derived male is indicated by a male (Mars) symbol. Scale bars: 1 mm (a), 10 μm (b), 50 μm (c).
