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Isolation and characterization of the human ovarian cell population for transplantation into an artificial ovary

机译：人体卵巢细胞群的分离和鉴定，用于移植到人工卵巢中

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To support survival and growth of follicles, the transplantable artificial ovary should mimic the original organ, offering a physical (3D matrix) and biological support (cells). In order to replicate the ovarian cell populations, the aim of this study is to assess the proportions of stromal and endothelial cells in the ovarian cortex. To this end, ovarian biopsies were obtained from six women (mean age: 49 years). The epithelial layer and medulla were carefully removed. The cortex was finely minced and enzymatically digested and the isolated cells were fixed. For cell characterization, immunostaining for CD31 (for endothelial cells) and inhibin-α (for granulosa cells) was performed. Positive cells in each staining were counted and the proportion of the different cell populations was estimated from the total number of isolated cells. Since there is no specific marker for ovarian stromal cells, we estimated the proportion of these cells by performing a vimentin immunostaining and subtracting the proportions of CD31- and inhibin-α-positive cells. Immunostaining showed that 84% of isolated cells were vimentin-positive. From this pool, 3% were endothelial cells and 1% granulosa cells. Consequently, the population of ovarian stromal cells was 80%. In conclusion, our findings show that stromal cells represent the larger population of cells in the human ovarian cortex. While this ensures follicle survival and development in a normal ovary, we believe that the low proportion of endothelial cells could have a negative impact on the angiogenesis in the artificial ovary after the first days of transplantation.
机译：为了支持卵泡的存活和生长，可移植的人工卵巢应模仿原始器官，提供物理（3D矩阵）和生物支持（细胞）。为了复制卵巢细胞群，本研究的目的是评估卵巢皮质中基质细胞和内皮细胞的比例。为此，从六名妇女（平均年龄：49岁）中获得了卵巢活检。小心去除上皮层和延髓。将皮层切碎并酶消化，将分离的细胞固定。为了进行细胞表征，对CD31（对于内皮细胞）和抑制素-α（对于颗粒细胞）进行了免疫染色。对每种染色中的阳性细胞进行计数，并从分离的细胞总数中估计不同细胞群的比例。由于没有针对卵巢基质细胞的特异性标记，我们通过进行波形蛋白免疫染色并减去CD31-和抑制素-α阳性细胞的比例来估计这些细胞的比例。免疫染色显示84％的分离细胞为波形蛋白阳性。在该池中，内皮细胞占3％，颗粒细胞占1％。因此，卵巢基质细胞的数量为80％。总之，我们的发现表明基质细胞代表了人类卵巢皮质中较大的细胞群。虽然这可以确保正常卵巢中的卵泡存活和发育，但我们认为移植后第一天内皮细胞的比例低可能会对人造卵巢中的血管生成产生负面影响。

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