A novel two-step strategy for in vitro culture of early-stage ovarian follicles in the mouse.
A novel two-step strategy for in vitro culture of early-stage ovarian follicles in the mouse.
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DOI：
10.1016/j.fertnstert.2009.10.027
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发表时间：
2010-05-15
影响因子：
6.7
通讯作者：
Woodruff TK
中科院分区：
文献类型：
--
作者：
Jin SY;Lei L;Shikanov A;Shea LD;Woodruff TK
开发一种体外策略，以支持早期卵泡的生长，并产生能够受精的成熟卵母细胞。取8日龄小鼠的全卵巢培养4d，分离次级卵泡，在三维海藻酸盐或纤维蛋白-海藻酸盐水凝胶基质中培养12d。大学附属实验室。小鼠没有对卵泡发育、类固醇激素产生和卵母细胞成熟率、卵母细胞受精和胚胎形成进行组织学评估。8日龄小鼠卵巢培养4d后，卵泡数量由原始卵泡和初级卵泡向次级卵泡过渡，与12日龄卵巢相似。培养12天的分离次级卵泡在FA-水凝胶基质中的卵母细胞直径增加、腔形成和膜细胞分化的频率均高于藻酸盐基质(P<；0.05)。类固醇激素分泌模式与培养的卵泡形态和细胞分化的变化相一致。与藻酸盐卵泡培养的卵母细胞相比，以FA为基础的卵泡培养获得的卵母细胞更多地发展到中期I(MI)和中期II(MII)，并且可以受精并分裂成两细胞胚胎(P<；0.05)。器官培养+FA水凝胶卵泡培养策略获得了极高的卵母细胞进展率(88±8.7%)和2-细胞胚胎形成率(54±4%)。早期卵泡的全卵巢培养和随后的FA水凝胶体外卵泡培养相结合的策略可以产生高比例的卵母细胞，并可能为面临威胁生育能力的疾病或治疗的妇女和青春期前女孩提供新的选择。
To develop an in vitro strategy to support the growth of early-stage follicles and produce mature oocytes competent for fertilization. Whole ovaries from 8-day-old mice were cultured for 4 days, then secondary follicles were isolated and cultured for 12 days in a 3-dimensional (3-D) alginate or fibrin-alginate hydrogel matrix. University-affiliated laboratory. Mice None Histologic evaluation of follicle development, steroid hormone production, and rates of oocyte maturation, oocyte fertilization, and embryo formation. Culture of 8-day-old mouse ovaries for 4 days resulted in transition of the follicle population from primordial and primary follicles to secondary follicles, similar to that seen in a 12-day-old ovary. Isolated secondary follicles cultured for 12 days showed larger increases in oocyte diameter and more frequent antrum formation and theca cell differentiation in the FA-hydrogel matrix compared with the alginate matrix (P<0.05). Steroid hormone secretion patterns were consistent with the changes in follicle morphology and cell differentiation observed in the cultured follicles. Compared with oocytes from alginate follicle cultures, a greater number of oocytes retrieved from the FA-based follicle cultures progressed to metaphase I (MI), reached metaphase II (MII) and could be fertilized and cleaved to two-cell embryos (P<0.05). The organ culture plus FA-hydrogel follicle culture strategy produced a very high rate of oocyte progression to MII (88 ± 8.7%) and formation of 2-cell embryos (54 ± 4%). A strategy combining whole ovary culture of early-stage follicles and subsequent FA hydrogel in vitro follicle culture produced a high percentage of oocytes competent for fertilization and may provide new options for fertility preservation in women and prepubertal girls facing fertility-threatening diseases or treatments.
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