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Abstract：
OBJECTIVE: Theca interna cells (TICs) are an indispensable cell source for ovarian follicle development and steroidogenesis. Recent studies have identified theca stem cells (TSCs) in both humans and animals. Interestingly, TSCs express mesenchymal stem cell (MSC)-related markers and can differentiate into mesenchymal lineages. MSCs are promising for tissue engineering and regenerative medicine due to their self-renewal and differentiation abilities. Therefore, this study investigated the potential origin of TICs from MSCs.
METHODS: Whole ovaries from postmenopausal organ donors were obtained, and their cortex was cryopreserved prior to the isolation of stromal cells. These isolated cells were differentiated in vitro to TICs using cell media enriched with various growth factors and hormones. Immunocytochemistry, an enzyme-linked immunosorbent assay, flow cytometry, and reverse transcription-quantitative polymerase chain were employed at different timepoints. Data were analyzed using one-way ANOVA.
RESULTS: Immunocytochemistry showed an increase in TIC markers from day 0 to day 8 and a significant rise in MSC-like markers on day 2. This corresponds with rising androstenedione levels from day 2 to day 13. Flow cytometry identified a decreasing MSC-like cell population from day 2 onwards. The CD13+ cell population and its gene expression increased significantly over time. NGFR and PDGFRA expression was induced on days 0 and 2, respectively, compared to day 13.
CONCLUSIONS: This study offers insights into MSC-like cells as the potential origin of TICs. Differentiating TICs from these widely accessible MSCs holds potential significance for toxicity studies and investigating TIC-related disorders like polycystic ovary syndrome (PCOS).
METHODS: Whole ovaries from postmenopausal organ donors were obtained, and their cortex was cryopreserved prior to the isolation of stromal cells. These isolated cells were differentiated in vitro to TICs using cell media enriched with various growth factors and hormones. Immunocytochemistry, an enzyme-linked immunosorbent assay, flow cytometry, and reverse transcription-quantitative polymerase chain were employed at different timepoints. Data were analyzed using one-way ANOVA.
RESULTS: Immunocytochemistry showed an increase in TIC markers from day 0 to day 8 and a significant rise in MSC-like markers on day 2. This corresponds with rising androstenedione levels from day 2 to day 13. Flow cytometry identified a decreasing MSC-like cell population from day 2 onwards. The CD13+ cell population and its gene expression increased significantly over time. NGFR and PDGFRA expression was induced on days 0 and 2, respectively, compared to day 13.
CONCLUSIONS: This study offers insights into MSC-like cells as the potential origin of TICs. Differentiating TICs from these widely accessible MSCs holds potential significance for toxicity studies and investigating TIC-related disorders like polycystic ovary syndrome (PCOS).
摘要：
目的：卵泡内膜细胞（TIC）是卵巢卵泡发育和类固醇生成不可或缺的细胞来源。最近的研究已经确定了人类和动物的卵泡膜干细胞（TSCs）。有趣的是,TSCs表达间充质干细胞（MSC）相关标记，并可分化成间充质谱系。由于MSCs具有自我更新和分化能力，因此有望用于组织工程和再生医学。因此,这项研究调查了来自MSCs的TIC的潜在来源。
方法：从绝经后器官捐献者获得全卵巢，在分离基质细胞之前，他们的皮质被冷冻保存。使用富含各种生长因子和激素的细胞培养基将这些分离的细胞体外分化为TIC。免疫细胞化学,酶联免疫吸附试验,流式细胞术,在不同的时间点使用逆转录定量聚合酶链。使用单向ANOVA分析数据。
结果：免疫细胞化学显示，从第0天到第8天，TIC标志物增加，第2天，MSC样标志物显著增加。这对应于从第2天到第13天升高的雄烯二酮水平。流式细胞术鉴定从第2天开始减少的MSC样细胞群。CD13+细胞群及其基因表达随时间显著增加。分别在第0天和第2天诱导NGFR和PDGFRA表达，与第13天相比。
结论：本研究为MSC样细胞作为TIC的潜在起源提供了见解。从这些可广泛获得的MSC中区分TIC对于毒性研究和研究TIC相关疾病如多囊卵巢综合征（PCOS）具有潜在的意义。
方法：从绝经后器官捐献者获得全卵巢，在分离基质细胞之前，他们的皮质被冷冻保存。使用富含各种生长因子和激素的细胞培养基将这些分离的细胞体外分化为TIC。免疫细胞化学,酶联免疫吸附试验,流式细胞术,在不同的时间点使用逆转录定量聚合酶链。使用单向ANOVA分析数据。
结果：免疫细胞化学显示，从第0天到第8天，TIC标志物增加，第2天，MSC样标志物显著增加。这对应于从第2天到第13天升高的雄烯二酮水平。流式细胞术鉴定从第2天开始减少的MSC样细胞群。CD13+细胞群及其基因表达随时间显著增加。分别在第0天和第2天诱导NGFR和PDGFRA表达，与第13天相比。
结论：本研究为MSC样细胞作为TIC的潜在起源提供了见解。从这些可广泛获得的MSC中区分TIC对于毒性研究和研究TIC相关疾病如多囊卵巢综合征（PCOS）具有潜在的意义。
