人卵泡膜细胞的体外发生、分化及其用于卵泡发育微环境体外重构的机制研究

卵泡膜细胞是卵泡发育微环境重要组成部分，体外重构卵泡微环境是实现人卵泡体外发育获得成熟卵母细胞的重要手段。既往哺乳动物研究发现，卵母细胞和颗粒细旁分泌调控卵泡膜细胞发生和分化，然而，人卵泡膜细胞的体内发生和分化旁分泌调控机制尚不清楚，难以体外有效建立卵泡发育微环境。本研究首先在组织和单细胞水平，通过单细胞转录组学研究技术建立人卵泡膜细胞体内分化过程中不同关键阶段的卵泡基因表达谱和膜细胞基因表达谱，筛选差异表达基因，进而利用膜细胞系进行功能验证；然后，在体外验证关键基因在人卵巢间质细胞向卵泡膜细胞分化进程中的作用，建立诱导卵泡膜细胞体外分化的实验体系，进而利用组织工程手段，将其与人卵泡共培养，优化窦前卵泡体外培养体系。旨在揭示人卵泡膜细胞体内、体外分化各阶段特异性的基因表达规律和关键调控蛋白，重构人卵泡体外发育微环境，填补人卵泡膜细胞发生机制研究的理论空白。

Theca cells are an important part of the folliculargenesis microenvironment. Efficient in vitro reconstruction of follicular microenvironment is the key point to promote human follicles in vitro growth to product mature oocytes. Mammals research found that Theca cells are differentiatied from interstitial cells under the control of oocyte and granulosa cells, however, human associated mechanisms has not been exactly in-depth studied, resulting in the failure of in vitro folliculargenesisi system.Therefore, in the first part of this study, we will investigate the transcriptome spectrum of human theca cell during differentiation in vivo through single cell techiniques and to screening the key factors regulating theca cell differentation; In the second part, we will explore the experimental system for theca cell differentation in vitro, and then reconstruct folliculargenesis microenvironment by using human theca cells. The aim is to explore the stage specific gene expression pattern of key regulatory proteins during human theca cell differentiation, and then to establish in vitro culture system for human preantral follicles. These results will contribute to breaking the bottleneck limiting human preantral follicles development in vitro, and making one step towards the fertility preservation for cancer patients to obtain their desired baby.