Lee Yeon-Mi, Kim Tae-Ho, Lee Jeong-Hyeon, Lee Won-Jae, Jeon Ryoung-Hoon, Jang Si-Jung, Ock Sun-A, Lee Sung-Lim, Park Bong-Wook, Rho Gyu-Jin
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Suwon, 441-706, Republic of Korea.
J Ovarian Res. 2016 Apr 12;9:24. doi: 10.1186/s13048-016-0233-z.
Recent findings have revealed that the female gonad may have regenerative activity with having germ line stem cells in juveniles and adults. Application of these germ line stem cells could be an alternative therapy for reproductive disorders in regenerative medicine.
To enhance the potency of differentiation into oocyte-like cells (OLCs) and folliculogenesis, we overexpressed Oct4 in ovarian stem/stromal cell (OvSCs) and examined the cellular properties related to stemness and self-renewal ability and finally demonstrated the ability of in vitro differentiation and folliculogenesis.
Ovarian cortex included putative stem cells in terms of AP activity, cell cycle status, cell proliferation, expression of mesenchymal lineage surface markers and pluripotent transcriptional markers. Further, Oct4 transfected OvSCs (Oct4-OvSCs) were enhanced their AP activity and cell proliferation compared to OvSCs. The potential on in vitro differentiation into OLCs and in vivo folliculogenesis was also evaluated in OvSCs and Oct4-OvSCs, respectively. Oct4-OvSCs possessed higher oogenesis potential in vitro than OvSCs, in terms of expression of germ cell markers by RT-PCR and the number of OLCs. When OvSCs and Oct4-OvSCs were xeno-transplanted into infertile mice ovaries, the OvSCs transplantation induced new primary follicle formation and hormonal levels of estradiol and FSH remained similar to that of normal mice. However, Oct4-OvSCs possessed higher ability for folliculogenesis based on inducing developing follicles with thecal layer and granulosa cells and more similar estradiol level to normal mice.
These findings demonstrated that putative stem cells were present in ovarian cortex and exhibited differentiation ability into OLCs and folliculogenesis in vivo, and Oct4-overexpression enhanced these ability, suggesting their cellular models based on gene therapy in understanding the mechanisms of oogenesis and folliculogenesis, and finally in view of reproductive cell therapy.
最近的研究结果表明，雌性性腺在幼年和成年期可能具有生殖活性，含有生殖系干细胞。这些生殖系干细胞的应用可能成为再生医学中治疗生殖障碍的一种替代疗法。
为了增强向卵母细胞样细胞（OLCs）分化和卵泡形成的能力，我们在卵巢干/基质细胞（OvSCs）中过表达Oct4，并检测与干性和自我更新能力相关的细胞特性，最终证明了体外分化和卵泡形成的能力。
从碱性磷酸酶（AP）活性、细胞周期状态、细胞增殖、间充质谱系表面标志物和多能转录标志物的表达来看，卵巢皮质包含推定的干细胞。此外，与OvSCs相比，Oct4转染的OvSCs（Oct4-OvSCs）的AP活性和细胞增殖增强。还分别在OvSCs和Oct4-OvSCs中评估了体外分化为OLCs和体内卵泡形成的潜力。就逆转录聚合酶链反应（RT-PCR）检测的生殖细胞标志物表达和OLCs数量而言，Oct4-OvSCs在体外具有比OvSCs更高的卵子发生潜力。当将OvSCs和Oct4-OvSCs异种移植到不育小鼠卵巢中时，OvSCs移植诱导了新的初级卵泡形成，雌二醇和促卵泡生成素（FSH）的激素水平与正常小鼠相似。然而，基于诱导形成具有卵泡膜层和颗粒细胞的发育卵泡以及与正常小鼠更相似的雌二醇水平，Oct4-OvSCs具有更高的卵泡形成能力。
这些发现表明，推定的干细胞存在于卵巢皮质中，并在体内表现出向OLCs分化和卵泡形成的能力，Oct4过表达增强了这些能力，这表明它们是基于基因治疗的细胞模型，有助于理解卵子发生和卵泡形成的机制，最终有望用于生殖细胞治疗。
