Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity.

Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity.
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DOI:
10.1038/s41598-018-25116-1
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发表时间:
2018-05-03
期刊:
影响因子:
4.6
通讯作者:
Telfer EE
Telfer EE
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Clarkson YL;McLaughlin M;Waterfall M;Dunlop CE;Skehel PA;Anderson RA;Telfer EE

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成年哺乳动物卵巢中存在一群具有种系发育潜力的推定干细胞(卵原干细胞:OSC),这一点在分离方法和体外转化潜力方面存在争议,特别是在细胞分选基于 DEAD box 多肽 4 (DDX4) 表达的情况下。这项研究描述了一种针对成年女性卵巢的精细组织解离/荧光激活细胞分选 (FACS) 方案,该方案可提高细胞活力和假定 OSC 的产量。结合 DDX4 和乙醛脱氢酶 1 (ALDH1) 双重检测的 FACS 技术证明存在两个具有 ALDH1 活性的小 DDX4 阳性细胞亚群(直径约 7μm),其特征在于差异剪接的 DDX4 转录物和 DAZL(生殖细胞分化的主要调节因子)的表达。这些可能表明祖细胞群的分化阶段,并为之前报道的表达差异提供可能的解释。这些发现为进一步表征这些细胞并探索其潜在的生理作用和治疗应用提供了坚实的基础。
The existence of a population of putative stem cells with germline developmental potential (oogonial stem cells: OSCs) in the adult mammalian ovary has been marked by controversy over isolation methodology and potential for in-vitro transformation, particularly where cell sorting has been based on expression of DEAD box polypeptide 4 (DDX4). This study describes a refined tissue dissociation/fluorescence-activated cell sorting (FACS) protocol for the ovaries of adult women which results in increased cell viability and yield of putative OSCs. A FACS technique incorporating dual-detection of DDX4 with aldehyde dehydrogenase 1 (ALDH1) demonstrates the existence of two sub-populations of small DDX4-positive cells (approx. 7 µm diameter) with ALDH1 activity, distinguished by expression of differentially spliced DDX4 transcripts and of DAZL, a major regulator of germ cell differentiation. These may indicate stages of differentiation from a progenitor population and provide a likely explanation for the expression disparities reported previously. These findings provide a robust basis for the further characterisation of these cells, and exploration of their potential physiological roles and therapeutic application.
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