Isolated primate primordial follicles require a rigid physical environment to survive and grow in vitro
Isolated primate primordial follicles require a rigid physical environment to survive and grow in vitro
复制标题
DOI：
10.1093/humrep/der468
复制
发表时间：
2012-06-01
影响因子：
6.1
通讯作者：
Woodruff, T. K.
中科院分区：
文献类型：
--
作者：
Hornick, J. E.;Duncan, F. E.;Woodruff, T. K.
体外卵泡生长是一种很有前途的生育力保存策略，其中卵巢卵泡被培养以产生成熟和具有受精能力的卵母细胞。然而，在灵长类动物中，从原始卵泡和初级卵泡开始的体外卵泡生长成功有限，因为尚未建立足够的分离方法和培养策略。了解如何利用原始卵泡保存生育能力具有重要意义，因为这些卵泡在卵巢中最丰富，在所有雌性动物中都有发现，并且对冷冻保存和化疗具有相当的抵抗力。在灵长类动物卵巢中，原始卵泡集中在富含胶原蛋白的卵巢皮质附近。为了获得这些卵泡，我们在酶消化之前分离卵巢皮质，并通过使用新型的双重过滤系统来富集原始卵泡浓度。为了验证体内刚性物理环境对存活最佳的假设，将原始卵泡在不同浓度的藻酸盐中培养长达6天。我们发现灵长类动物的卵巢组织在4 ℃下可以维持24小时而不损害组织或卵泡的健康。从每个卵巢中分离出数百个完整且有活力的原始卵泡，与动物年龄无关。与0.5藻酸盐相比，当卵泡在2藻酸盐中培养时，卵泡存活和形态更理想。通过使用生物材料模拟刚性卵巢环境，我们建立了支持原始卵泡培养的条件。这些结果奠定了基础，研究在一个可控的环境中的原始卵泡的基础生物学和使用原始卵泡的生育力保存方法。
In vitro follicle growth is a promising fertility preservation strategy in which ovarian follicles are cultured to produce mature and fertilization-competent oocytes. However, in primates, there has been limited success with in vitro follicle growth starting from primordial and primary follicles because adequate isolation methods and culture strategies have not been established. Understanding how to use primordial follicles for fertility preservation has significant implications because these follicles are the most abundant in the ovary, are found in all females and are fairly resistant to cryopreservation and chemotherapeutics.In the primate ovary, primordial follicles are concentrated near the collagen-rich ovarian cortex. To obtain these follicles, we separated the ovarian cortex prior to enzymatic digestion and enriched the primordial follicle concentration by using a novel double filtration system. To test the hypothesis that a rigid physical environment, as found in vivo, is optimal for survival, primordial follicles were cultured in different concentrations of alginate for up to 6 days. Follicle survival and morphology were monitored throughout the culture.We found that primate ovarian tissue can be maintained for up to 24 h at 4C without compromising tissue or follicle health. Hundreds of intact and viable primordial follicles were isolated from each ovary independent of animal age. Follicle survival and morphology were more optimal when follicles were cultured in 2 alginate compared with 0.5 alginate.By mimicking the rigid ovarian environment through the use of biomaterials, we have established conditions that support primordial follicle culture. These results lay the foundations for studying the basic biology of primordial follicles in a controlled environment and for using primordial follicles for fertility preservation methods.