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

• Published in: Scientific reports Vol. 8; no. 1; pp. 6953 - 11
• Main Authors: Clarkson, Yvonne L, McLaughlin, Marie, Waterfall, Martin, Dunlop, Cheryl E, Skehel, Paul A, Anderson, Richard A, Telfer, Evelyn E
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 03.05.2018; Nature Publishing Group UK; Nature Portfolio
• Subjects: Aldehyde dehydrogenase; Aldehyde Dehydrogenase 1 Family; Cell Separation - methods; Cells, Cultured; DEAD-box RNA Helicases - analysis; DEAD-box RNA Helicases - genetics; Dehydrogenases; Female; Flow cytometry; Flow Cytometry - methods; Gene Expression; Humans; Isoenzymes - analysis; Oogonial Stem Cells - cytology; Oogonial Stem Cells - metabolism; Ovaries; Ovary - cytology; Ovary - metabolism; Retinal Dehydrogenase - analysis; Stem cells; Young Adult
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-25116-1

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.