School of Medicine, Department of Obstetrics and Gynecology, Reproductive Endocrinology and Infertility and Assisted Reproductive Technology (ART), Kocaeli University, Unit. Izmit, 41380, Kocaeli, Turkey.
Graduate School of Health Sciences, Department of Stem Cell, Center for Stem Cells and Gene Therapies Research and Practice, Kocaeli University, Izmit, 41380, Kocaeli, Turkey.
Stem Cell Rev Rep. 2019 Aug;15(4):558-573. doi: 10.1007/s12015-019-09892-5.
Premature ovarian insufficiency (POI), a fertility disorder affecting women under 40 years of age, is characterized by early loss of ovarian function. This study was aimed to maintain ovarian function in POI animal models by mesenchymal stem cells (MSCs) transplantation with/without the supplementation of platelet-rich plasma (PRP). Adipose tissue-derived MSCs were isolated from inbred rats (Fisher-344), and constitutive expression of both VEGF and GFP were maintained by transfection with plasmids, pVEGF and pGFP-N. PRP was derived from the blood of healthy untreated rats. A total of 60 rats were divided into 5 groups of 12 rats in each. First group was kept as untreated-control (Control), and POI model was induced in Fisher-344 rats by cyclophosphamide in the next four groups. Second group was kept as sham-operated-control (Sham). MSC, PRP and MSC+ PRP-treated groups were transplanted following the validation of POI model in rats. After 2 months following the transplantation, anti-mullerian-hormone (AMH) and oestradiol (E) blood levels were measured. Follicles were evaluated after hematoxylin-eosin staining, and the immunofluorescence staining and gene expression analyses were performed to show the ovarian regeneration. The follicular count was improved after MSC- and MSC + PRP-treatment to 63% of Control-group and significantly higher than that in Sham-group, but a significant increase was not observed in PRP-group. Higher AMH and E levels were measured in MSC + PRP than in Sham-group, and CXCL12, BMP-4, TGF-β and IGF-1 expressions were also increased. This study showed MSCs +/-PRP transplantation after POI supports recovery of the follicular count and function. For ovarian recovery, a single administration of PRP was found not sufficient. Although MSC treatment increased follicular regeneration, better results were obtained in the co-transplantation of MSCs and PRP. These results might be promising for follicular regeneration in POI patients.
卵巢早衰（POI）是一种影响 40 岁以下女性生育能力的疾病，其特征是卵巢功能过早丧失。本研究旨在通过间充质干细胞（MSCs）移植并用/不用富含血小板的血浆（PRP）补充来维持 POI 动物模型中的卵巢功能。脂肪组织来源的 MSCs 从近交系大鼠（Fisher-344）中分离出来，并通过转染质粒 pVEGF 和 pGFP-N 维持 VEGF 和 GFP 的组成型表达。PRP 从健康未治疗大鼠的血液中提取。共有 60 只大鼠分为 5 组，每组 12 只。第一组作为未处理对照组（Control），下一组 Fisher-344 大鼠用环磷酰胺诱导 POI 模型。第二组作为假手术对照组（Sham）。在大鼠 POI 模型验证后，对 MSC、PRP 和 MSC+PRP 治疗组进行移植。移植后 2 个月，测量抗苗勒管激素（AMH）和雌二醇（E）的血液水平。用苏木精-伊红染色评估卵泡，进行免疫荧光染色和基因表达分析以显示卵巢再生。MSC 和 MSC+PRP 治疗后卵泡计数提高到 Control 组的 63%，明显高于 Sham 组，但 PRP 组未见明显增加。MSC+PRP 组的 AMH 和 E 水平高于 Sham 组，CXCL12、BMP-4、TGF-β和 IGF-1 的表达也增加。本研究表明，POI 后 MSC +/-PRP 移植支持卵泡计数和功能的恢复。对于卵巢恢复，单次 PRP 给药发现不够。虽然 MSC 治疗增加了卵泡再生，但 MSC 和 PRP 共移植的效果更好。这些结果可能对 POI 患者的卵泡再生有希望。
