Tsai Te-Sha, Johnson Jacqueline, White Yvonne, John Justin C
Centre for Genetic Diseases, Hudson Institute of Medical Research, Clayton, Victoria 3168, Australia.
Centre for Genetic Diseases, Department of Molecular and Translational Science, Monash University, Clayton, Victoria 3168, Australia.
Oncotarget. 2017 Jun 28;8(38):63484-63505. doi: 10.18632/oncotarget.18833. eCollection 2017 Sep 8.
Female-factor infertility can be caused by poor oocyte quality and depleted ovarian reserves. Egg precursor cells (EPCs), isolated from the ovarian cortex, have the potential to be used to overcome female infertility. We aimed to define the origins of EPCs by analyzing their gene expression profiles and mtDNA content using a mini-pig model. We characterized FAC-sorted DDX4-derived porcine EPCs by performing RNA-sequencing and determined that they utilize pathways important for cell cycle and proliferation, which supports the existence of adult mitotically active oogonial cells. Expression of the pluripotent markers and , and the primitive germ cell markers and were not detected. However, and were expressed, as were the primitive germ cell markers , and . Moreover, porcine EPCs expressed self-renewal and proliferation markers including , , , , , , , and . The presence of , , and were not detected, indicating that porcine EPCs are not indicative of mature primordial oocytes. We performed mitochondrial DNA Next Generation Sequencing and determined that one mtDNA variant harbored by EPCs was present in oocytes, preimplantation embryos and somatic tissues over three generations in our mini-pig model indicating the potential germline origin of EPCs.
女性因素不孕症可能由卵母细胞质量差和卵巢储备耗竭引起。从卵巢皮质分离出的卵母细胞前体细胞(EPCs)有潜力用于克服女性不孕症。我们旨在通过使用小型猪模型分析其基因表达谱和线粒体DNA含量来确定EPCs的起源。我们通过进行RNA测序对荧光激活细胞分选(FAC)筛选出的源自DDX4的猪EPCs进行了表征,并确定它们利用对细胞周期和增殖重要的途径,这支持了成年有丝分裂活跃的卵原细胞的存在。未检测到多能性标志物和,以及原始生殖细胞标志物和的表达。然而,和被表达,原始生殖细胞标志物、和也被表达。此外,猪EPCs表达自我更新和增殖标志物,包括、、、、、、、和。未检测到、、和的存在,表明猪EPCs不代表成熟的原始卵母细胞。我们进行了线粒体DNA下一代测序,并确定在我们的小型猪模型中,EPCs携带的一种线粒体DNA变体在三代的卵母细胞、植入前胚胎和体细胞组织中都存在,这表明EPCs可能起源于生殖系。