Abstract:
Primordial germ cells (PGCs) are germline-restricted embryonic cells that form the functional gametes of the adult animal. The use of avian PGCs in biobanking and producing genetically modified birds has driven research on the in vitro propagation and manipulation of these embryonic cells. In avian species, PGCs are hypothesized to be sexually undetermined at an early embryonic stage and undergo differentiation into an oocyte or spermatogonial fate dictated by extrinsic factors present in the gonad. However, chicken male and female PGCs require different culture conditions, suggesting that there are sex-specific differences, even at early stages. To understand potential differences between male and female chicken PGCs during migratory stages, we studied the transcriptomes of circulatory stage male and female PGCs propagated in a serum-free medium. We found that in vitro cultured PGCs were transcriptionally similar to their in ovo counterparts, with differences in cell proliferation pathways. Our analysis also revealed sex-specific transcriptome differences between male and female cultured PGCs, with notable differences in Smad7 and NCAM2 expression. A comparison of chicken PGCs with pluripotent and somatic cell types identified a set of genes that are exclusive to germ cells, enriched in the germplasm, and associated with germ cell development.
摘要:
原始生殖细胞(PGCs)是种系限制的胚胎细胞,其形成成年动物的功能配子。禽类PGC在生物捕获和生产遗传修饰的禽类中的使用推动了对这些胚胎细胞的体外繁殖和操作的研究。在鸟类中,假设PGCs在早期胚胎阶段是性不确定的,并经历了由性腺中存在的外在因素决定的卵母细胞或精原命运。然而,鸡雄性和雌性PGCs需要不同的培养条件,这表明存在性别差异,即使在早期阶段。为了了解迁徙阶段雄性和雌性鸡PGCs之间的潜在差异,我们研究了在无血清培养基中繁殖的循环期雄性和雌性PGCs的转录组。我们发现体外培养的PGCs在转录上与卵内的对应物相似,细胞增殖途径的差异。我们的分析还揭示了男性和女性培养的PGCs之间的性别特异性转录组差异,Smad7和NCAM2表达存在显著差异。将鸡PGCs与多能细胞和体细胞类型进行比较,确定了一组生殖细胞独有的基因,丰富的种质,与生殖细胞发育有关.