Transplantation of ovarian granulosa‑like cells derived from human induced pluripotent stem cells for the treatment of murine premature ovarian failure.
Transplantation of ovarian granulosa‑like cells derived from human induced pluripotent stem cells for the treatment of murine premature ovarian failure.
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移植源自人诱导多能干细胞的卵巢颗粒样细胞治疗小鼠卵巢早衰
DOI：
10.3892/mmr.2016.5191
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发表时间：
2016-06
影响因子：
3.4
通讯作者：
Zheng J
中科院分区：
文献类型：
--
作者：
Liu T;Li Q;Wang S;Chen C;Zheng J
卵巢早衰（POF）是女性不孕的常见原因，目前还没有理想的治疗方法或药物。此外，卵巢颗粒细胞（OGC）的凋亡是卵巢储备和功能下降的重要机制。在本研究中，几种细胞生长因子和激素诱导人诱导多能干细胞（iPSCs）分化为卵巢颗粒样细胞（OGLCs）在体外。免疫组织化学染色显示诱导后12天，iPSCs来源的OGLC强烈表达颗粒细胞标志物，包括抗苗勒管激素、BMP 1 α、BMP 1 β和促卵泡激素受体，但不表达干细胞标志物，包括八聚体结合转录因子4、SRY（性别决定区Y）-box 2、Nanog和阶段特异性胚胎抗原-4。此外，通过环磷酰胺治疗产生POF小鼠模型。随后，将iPSC衍生的OGLC体内移植到POF小鼠中（OGLCs-iPSC-POF组）。结果表明，与对照组（磷酸盐缓冲液处理组）相比，OGLCs的生长状态明显改善，OGLCs-iPSCs-POF组卵巢组织中可检测到成熟卵泡。免疫组织化学染色显示，iPSC衍生的OGLC移植到POF小鼠中不仅在小鼠卵巢组织中表现出显著的生长，而且还强烈表达OGC标记物。此外，酶联免疫吸附测定表明，外周血样品中的激素雌二醇的水平显着增强后，iPSC衍生的OGLC移植到POF小鼠。此外，与对照组相比，OGLCs-iPSCs-POF组中的卵巢组织重量显著更高，并且与对照小鼠相比，OGLCs-iPSCs-POF小鼠中的闭锁卵泡数量显著减少。这些结果表明，来源于人iPSC的OGLC不仅可以有效地增强OGC生长和修复受损的卵巢组织，而且还可以维持卵巢组织生态位，促进POF小鼠模型中的卵泡发育和成熟。
Premature ovarian failure (POF) is a common cause of female infertility, for which there are currently no ideal treatments or medications. Furthermore, apoptosis of ovarian granulosa cells (OGCs) is an important mechanism underlying the decline in ovarian reserve and function. In the present study, several cellular growth factors and hormones were used to induce the differentiation of human induced pluripotent stem cells (iPSCs) into ovarian granulosa-like cells (OGLCs) in vitro. Immunohistochemical staining demonstrated that OGLCs derived from iPSCs strongly expressed granulosa cell markers, including anti-Müllerian hormone, inhibin α, inhibin β and follicle-stimulating hormone receptor, but did not express stem cell markers, including octamer-binding transcription factor 4, SRY (sex determining region Y)-box 2, Nanog and stage-specific embryonic antigen-4 12 days post-induction. In addition, a mouse model of POF was generated by cyclophosphamide treatment. Subsequently, iPSC-derived OGLCs were transplanted into the POF mice (OGLCs-iPSCs-POF group) in vivo. Results indicated that, compared with the control group (POF mice treated with phosphate-buffered saline), the growth state of OGLCs was markedly improved, and mature follicles could be detected in the ovarian tissue of the OGLCs-iPSCs-POF group. Immunohistochemical staining demonstrated that iPSC-derived OGLCs transplanted into POF mice not only exhibited substantial growth in murine ovarian tissues, but also strongly expressed OGC markers. Furthermore, enzyme-linked immunosorbent assays indicated that the levels of the hormone estradiol in peripheral blood samples were significantly enhanced following transplantation of iPSC-derived OGLCs into POF mice. Furthermore, ovarian tissue weight was significantly higher in the OGLCs-iPSCs-POF group compared with in the control group, and the number of atretic follicles in OGLCs-iPSCs-POF mice was significantly reduced, as compared with in the control mice. These results suggest that OGLCs derived from human iPSCs may not only effectively enhance OGC growth and repair damaged ovarian tissue, but may also maintain the ovarian tissue niche, promoting follicular development and maturation in a mouse model of POF.