目的:尽管人类原始卵泡的体外发育取得了重大进展,这仍然是一个具有挑战性的方法,有很大的改进潜力。因此,本研究旨在研究人卵泡膜祖细胞(hTPCs)饲养层对嵌入人卵巢组织的原始卵泡发育的影响。
方法:使用钒酸衍生物双过氧(5-羟基-吡啶-2-羧酸)氧钒酸二钾(V)和试剂盒配体将冻融的卵巢组织片段活化24小时。将标本分为共培养和单一培养组,在有和没有hTPC饲养层的情况下培养6天,分别。之后,对卵泡进行计数和分类,并评估了激素水平和凋亡和卵泡发生相关基因的表达水平。
结果:两个培养组均显示出显着的卵泡生长(P<0.05)。然而,与其他组相比,共培养组的卵泡数量显着增加(P<0.05)。此外,ZP1、ZP2、ZP3、BMP-7、AMH、共培养组GDF9明显高于其他组(P<0.05),P53和CASP3的表达水平明显降低(P<0.05)。此外,雌二醇的浓度,黄体酮,睾丸激素,共培养组和雄烯二酮显著高于其他组(P<0.05)。
结论:本研究结果为hTPCs在人类原始卵泡的生长和发育中的直接作用提供了新的证据。然而,未来的研究需要说明潜在的机制。结果的示意图。根据我们的结果,ZP1,ZP2,ZP3和GDF9在卵母细胞中的表达,颗粒细胞中的AMH,共培养组卵泡膜细胞中的BMP4和BMP4显著高于单一培养和非培养组,而凋亡基因的表达(BAX,CASP3、P53)显著降低。此外,共培养组雌二醇水平显着增加,黄体酮,睾丸激素,与单一文化群体相比,其培养基中的雄烯二酮。
OBJECTIVE: Despite the significant advances in the in vitro development of human primordial follicles, it is still a challenging approach with great potential for improvements. Therefore, the present study aimed to investigate the effect of a feeder layer of human theca progenitor cells (hTPCs) on the development of primordial follicles embedded in human ovarian tissue.
METHODS: Fragments of frozen-thawed ovarian tissue were activated using the vanadate-derivative dipotassium bisperoxo (5-hydroxy-pyridine-2-carboxylic) oxovanadate (V) and kit ligand for 24 h. Then, the specimens were divided into the co-culture and mono-culture groups and were cultured with and without a hTPC feeder layer for 6 days, respectively. Afterward, the follicles were counted and classified, and the hormone levels and expression levels of apoptosis- and folliculogenesis-related genes were assessed.
RESULTS: Both culture groups showed significant follicle growth (P < 0.05). However, the co-culture group had a significantly higher number of growing follicles compared to the other group (P < 0.05). Moreover, the expression levels of ZP1, ZP2, ZP3, BMP-7, AMH, and GDF9 were significantly higher in the co-culture group compared to the other group (P < 0.05), while the expression levels of P53 and CASP3 were significantly lower (P < 0.05). Also, the concentrations of estradiol, progesterone, testosterone, and androstenedione were significantly higher in the co-culture group compared to the other group (P < 0.05).
CONCLUSIONS: The present study results provided novel evidence on the direct role of hTPCs in the growth and development of human primordial follicles. However, there is a need for future studies to illustrate the underlying mechanisms. Schematic summary of the results. According to our results, the expression of ZP1, ZP2, ZP3, and GDF9 in the oocytes, AMH in the granulosa cells, and BMP4 in the theca cells of the co-culture group were significantly higher than those of the mono-culture and non-culture groups, while the expression of apoptotic genes (BAX, CASP3, and P53) was significantly lower. Moreover, the co-culture group showed significantly increased levels of estradiol, progesterone, testosterone, and androstenedione in its culture media compared to the mono-culture groups.