Nakamura Yoshiaki, Usui Fumitake, Miyahara Daichi, Mori Takafumi, Ono Tamao, Takeda Kumiko, Nirasawa Keijiro, Kagami Hiroshi, Tagami Takahiro
Faculty of Agriculture, Shinshu University, Minamiminowa, Nagano 399-4598, Japan.
Reprod Fertil Dev. 2010;22(8):1237-46. doi: 10.1071/RD10056.
The unique accessibility of chicken primordial germ cells (PGCs) during early development provides the opportunity to combine the reproduction of live animals with genetic conservation. Male and female Gifujidori fowl (GJ) PGCs were collected from the blood of early embryos, and cryopreserved in liquid nitrogen for >6 months until transfer. Manipulated GJ embryos were cultured until hatching; fertility tests indicated that they had normal reproductive abilities. Embryos from two lines of White Leghorn (24HS, ST) were used as recipients for chimera production following blood removal. The concentration of PGCs in the early embryonic blood of 24HS was significantly higher than in ST (P < 0.05). Frozen-thawed GJ PGCs were microinjected into the bloodstream of same-sex recipients. Offspring originating from GJ PGCs in ST recipients were obtained with a higher efficiency than those originating from GJ PGCs in 24HS recipients (23.3% v. 3.1%). Additionally, GJ progeny were successfully regenerated by crossing germline chimeras of the ST group. In conclusion, the cryogenic preservation of PGCs from early chicken embryos was combined with the conservation of live animals.
鸡原始生殖细胞(PGCs)在早期发育过程中具有独特的可获取性,这为将活体动物繁殖与基因保存相结合提供了机会。从早期胚胎的血液中收集了雄性和雌性岐阜锦鸡(GJ)的PGCs,并在液氮中冷冻保存超过6个月,直至进行移植。对经过处理的GJ胚胎进行培养直至孵化;生育力测试表明它们具有正常的生殖能力。在去除血液后,使用来自两个白来航鸡品系(24HS、ST)的胚胎作为嵌合体生产的受体。24HS早期胚胎血液中PGCs的浓度显著高于ST(P < 0.05)。将冻融后的GJ PGCs显微注射到同性受体的血液中。与源自24HS受体中GJ PGCs的后代相比,源自ST受体中GJ PGCs的后代获得效率更高(23.3%对3.1%)。此外,通过使ST组的种系嵌合体杂交成功再生了GJ后代。总之,将早期鸡胚胎的PGCs低温保存与活体动物的保存相结合。