UNASSIGNED: Adult mammalian ovaries contain stem/progenitor cells necessary for folliculogenesis and ovulation-related tissue rupture repair. Theca cells are recruited and developed from progenitors during the folliculogenesis. Theca cell progenitors were not well defined. The aim of current study is to compare the potentials of four ovarian progenitors with defined markers (LY6A, EPCR, LGR5, and PDGFRA) to form steroidogenic theca cells in vitro. The location of the progenitors with defined makers was determined by immunohistochemistry and immunofluorescence staining of ovarian sections of adult mice. Different progenitor populations were purified by magnetic-activated cell sorting (MACS) and/or fluorescence-activated cell sorting (FACS) techniques from ovarian cell preparation and were tested for their abilities to generate steroidogenic theca cells in vitro. The cells were differentiated with a medium containing LH, ITS, and DHH agonist for 12 days. The results showed that EPCR+ and LGR5+ cells primarily distributed along the ovarian surface epithelium (OSE), while LY6A+ cells distributed in both the OSE and parenchyma. However, PDGFRA+ cells were exclusively located in interstitial compartment. When the progenitors were purified by these markers and differentiated in vitro, LY6A+ and PDGFRA+ cells formed steroidogenic cells expressing both CYP11A1 and CYP17A1 and primarily producing androgens, showing characteristics of theca-like cells, while LGR5+ cells generated steroidogenic cells devoid of CYP17A1 expression and androgen production, showing a characteristic of progesterone-producing cells (granulosa- or lutea-like cells). In conclusion, progenitors from both OSE and parenchyma of adult mice are capable of generating steroidogenic cells with different steroidogenic capacities, showing a possible lineage preference.
方法:卵巢祖细胞由上述4个先前报道的标记鉴定。通过对成年小鼠卵巢切片的免疫组织化学和免疫荧光染色来确定细胞的位置。通过磁性激活细胞分选(MACS)和/或荧光激活细胞分选(FACS)技术从卵巢细胞制剂中纯化不同的祖细胞群,并测试其体外生成类固醇性卵泡膜细胞的能力。细胞用含有LH的培养基分化,ITS和DHH激动剂持续12天。
结果:EPCR+和LGR5+细胞主要分布在卵巢表面上皮(OSE),而LY6A+细胞分布在OSE和薄壁组织中。然而,PDGFRA+细胞仅位于间质区室中。当通过这些标记纯化祖细胞并在体外分化时,LY6A+和PDGFRA+细胞形成了表达CYP11A1和CYP17A1并主要产生雄激素的类固醇细胞,表现出卵泡膜样细胞的特征,而LGR5+细胞产生的类固醇细胞缺乏CYP17A1表达和雄激素产生,表现出孕酮产生细胞(颗粒细胞或lutea样细胞)的特征。
结论:来自成年小鼠的OSE和薄壁组织的祖细胞能够产生具有不同类固醇生成能力的类固醇生成细胞,显示可能的血统偏好。