Many modern techniques employed to uncover the molecular fundamentals underlying biological processes require dissociated cells as their starting point/substrate. Investigations into ovarian endocrinology or folliculogenesis, therefore, necessitate robust protocols for dissociating the ovary into its constituent cell populations. While in the mouse, methods to obtain individual, mature follicles are well-established, the separation and isolation of single cells of all types from early mouse follicles, including somatic cells, has been more challenging. Herein we present two methods for the isolation of somatic cells in the ovary. These methods are suitable for a range of applications relating to the study of folliculogenesis and mouse ovarian development. First, an enzymatic dissociation utilising collagenase and a temporary, primary cell culture step using neonatal mouse ovaries which yields large quantities of granulosa cells from primordial, activating, and primary follicles. Second, a rapid papain dissociation resulting in a high viability single cell suspension of ovarian somatic cells in less than an hour, which can be applied from embryonic to adult ovarian samples. Collectively these protocols can be applied to a broad array of investigations with unique advantages and benefits pertaining to both.

中文翻译：

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从小鼠卵巢中提取体细胞群的两种替代方法

用于揭示生物过程的分子基础的许多现代技术需要解离的细胞作为它们的起点/底物。因此，对卵巢内分泌或卵泡发生的研究需要强有力的方案来将卵巢分离为其组成细胞群。虽然在小鼠中，获得单个成熟卵泡的方法已经成熟，但从早期小鼠卵泡（包括体细胞）中分离和分离所有类型的单细胞更具挑战性。在这里，我们提出了两种分离卵巢中体细胞的方法。这些方法适用于与研究卵泡发生和小鼠卵巢发育相关的一系列应用。首先，利用胶原酶的酶解和暂时的，使用新生小鼠卵巢进行原代细胞培养步骤，从原始卵泡、活化卵泡和初级卵泡中产生大量颗粒细胞。其次，木瓜蛋白酶的快速解离可在不到一小时的时间内产生高活力的卵巢体细胞单细胞悬浮液，可应用于从胚胎到成人卵巢样本。总的来说，这些协议可以应用于广泛的调查，具有与两者相关的独特优势和好处。