San Diego Zoo Institute for Conservation Research, Escondido, CA 92027, USA.
Biol Reprod. 2013 May 23;88(5):129. doi: 10.1095/biolreprod.112.105189. Print 2013 May.
As advanced reproductive technologies have become routine for domesticated species, they have begun to be applied in the field of endangered species conservation. For avian conservation, the most promising technology is the transfer of germ stem cells of exotic species to domestic hosts for the production of gametes. In this study, adult quail (model for exotic species) spermatogonial stem cells were xenogeneically transferred to stages 14-17 chicken host embryos. Fluorescent cellular dyes, quail-specific antibodies, and quail-specific quantitative PCR confirmed donor cell migration to and colonization of the host gonadal ridge. Donor-derived cells were observed by fluorescent microscopy in the caudal area as early as 2 h after injection, in the gonadal ridge at 4 h after injection, as well as in the gonads of stages 35-38 host embryos. Four of eight donor-derived cell flow cytometry-positive host gonads were confirmed by quantitative PCR using quail-specific primers. There was no statistically significant effect of host stage of injection, host gonad isolation stage, or host sex on the number of hosts positive for donor cells or the percent of donor-derived cells per positive gonad. Donor-derived cells isolated from stages 35-38 host gonads costained with the germ stem cell marker SSEA-1, indicating that the donor-derived cells have maintained stem cell-ness. This is the first study to suggest that it is feasible to rescue adult germ stem cells of deceased birds to prolong the reproductive lifespan of critically endangered species or genetically valuable individuals by transferring them to an embryonic chicken host.
随着先进的生殖技术在驯化物种中变得常规化,它们开始应用于濒危物种保护领域。对于鸟类保护而言,最有前途的技术是将外来物种的生殖干细胞转移到国内宿主中,以产生配子。在这项研究中,成年鹌鹑(外来物种模型)精原干细胞被异种移植到鸡胚的 14-17 期。荧光细胞染料、鹌鹑特异性抗体和鹌鹑特异性定量 PCR 证实了供体细胞向宿主生殖嵴的迁移和定植。注射后 2 小时即可在尾部区域通过荧光显微镜观察到供体细胞,4 小时后可在注射后的生殖嵴中观察到,在 35-38 期宿主胚胎的性腺中也可观察到。使用鹌鹑特异性引物的定量 PCR 证实了 8 个供体衍生细胞流式细胞术阳性的宿主性腺中的 4 个。宿主注射阶段、宿主性腺分离阶段或宿主性别对供体细胞阳性宿主的数量或每个阳性性腺中供体衍生细胞的百分比均无统计学显著影响。从 35-38 期宿主性腺中分离出的供体衍生细胞与生殖干细胞标记物 SSEA-1 共染色,表明供体衍生细胞保持了干细胞特性。这是第一项表明通过将成年生殖干细胞转移到鸡胚宿主中,可以拯救濒危物种或具有遗传价值的个体的成年生殖干细胞,从而延长其生殖寿命的研究。