Initiation of growth of baboon primordial follicles in vitro
Initiation of growth of baboon primordial follicles in vitro
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DOI：
10.1093/humrep/12.9.1993
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发表时间：
1997-09-01
影响因子：
6.1
通讯作者：
Fortune, JE
中科院分区：
文献类型：
--
作者：
Wandji, SA;Srsen, V;Fortune, JE
灵长类原始卵泡可以在无血清培养基中存活并开始体外生长的假设得到了验证。从妊娠晚期的狒狒胎儿获得卵巢皮质浅层，主要含有原始卵泡，并培养0，2，4，7，在添加胰岛素、转铁蛋白、硒、亚油酸和牛血清白蛋白（ITS+）的Waymouth MB 752/1培养基中培养10或20天后，皮质片的组织学检查显示，培养2天和4天后，原始卵泡总数分别减少了55%和76%（P < 0.01），这与总初级卵泡数持续增加5- 8倍有关（P < 0.01）从培养第2天开始，早期次级卵泡和次级卵泡总数也逐渐增加，除次级卵泡和卵母细胞外，所有卵泡类型的卵泡和卵母细胞的平均直径在整个培养过程中均逐渐增大（P < 0.01）。增殖细胞核抗原（PCNA），细胞增殖和生长的标志物，免疫组织化学定位显示，PCNA是一般情况下，在原始卵泡0天，但观察到培养后2或4天，在大多数生长卵泡的颗粒细胞和卵母细胞。比较用ITS+和10%胎牛血清（FBS）培养10或20天的皮质片，显示与FBS处理的皮质片相比，ITS+处理的皮质片中的原始卵泡数量减少更明显，初级卵泡、早期次级卵泡和次级卵泡更多这些结果表明，非人灵长类原始卵泡在无血清条件下可以在体外存活并发育到次级阶段。
Factors that cause some primordial follicles to enter the growth phase while the others remain quiescent are unknown, The hypothesis was tested that primate primordial follicles can survive and initiate growth in vitro in serum-free medium, Superficial pieces of ovarian cortex, containing mostly primordial follicles, were obtained from baboon fetuses during late gestation and cultured for 0, 2, 4, 7, 10 or 20 days in Waymouth MB 752/1 medium supplemented with insulin, transferrin, selenium, linoleic acid, and bovine serum albumin (ITS+), Histological examination of cortical pieces revealed that after 2 and 4 days in culture, the total number of primordial follicles had decreased by 55 and 76% (P < 0.01) respectively, relative to day 0 of culture, This was associated with a sustained, 5- to 8-fold increase in total primary follicles (P < 0.01) beginning on day 2 of culture, There was also a gradual increase in the total number of early secondary and secondary follicles, The average diameter of follicles and oocytes increased gradually throughout culture for all follicular categories (P < 0.01), except secondary follicles and oocytes. Immunohistochemical localization of proliferating cell nuclear antigen (PCNA), a marker for cell proliferation and growth, showed that PCNA was generally absent in primordial follicles on day 0, but was observed after 2 or 4 days in culture in both granulosa cells and oocytes of most growing follicles. Comparison of cortical pieces cultured for 10 or 20 days with ITS+ versus 10% fetal bovine serum (FBS) showed a more pronounced decrease in the numbers of primordial follicles and more primary, early secondary and secondary follicles in ITS+ compared to FBS-treated cortical pieces (P < 0.01 at 20 days), These results show that primordial follicles from non-human primates can survive and develop to the secondary stage in vitro in serum-free conditions.