Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity
Abstract 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 b...
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| Format: | Article |
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Nature Portfolio
2018-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-018-25116-1 |
| _version_ | 1818754246512738304 |
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| author | Yvonne L. Clarkson Marie McLaughlin Martin Waterfall Cheryl E. Dunlop Paul A. Skehel Richard A. Anderson Evelyn E. Telfer |
| author_facet | Yvonne L. Clarkson Marie McLaughlin Martin Waterfall Cheryl E. Dunlop Paul A. Skehel Richard A. Anderson Evelyn E. Telfer |
| author_sort | Yvonne L. Clarkson |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-18T05:20:12Z |
| format | Article |
| id | doaj.art-52dccacc26814c61baeefdbc37202808 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-18T05:20:12Z |
| publishDate | 2018-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-52dccacc26814c61baeefdbc372028082022-12-21T21:19:40ZengNature PortfolioScientific Reports2045-23222018-05-018111110.1038/s41598-018-25116-1Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activityYvonne L. Clarkson0Marie McLaughlin1Martin Waterfall2Cheryl E. Dunlop3Paul A. Skehel4Richard A. Anderson5Evelyn E. Telfer6Institute of Cell Biology and the Centre for Integrative Physiology, Hugh Robson Building, University of EdinburghInstitute of Cell Biology and the Centre for Integrative Physiology, Hugh Robson Building, University of EdinburghSchool of Biological Sciences, Ashworth Laboratories, the King’s Buildings, University of EdinburghInstitute of Cell Biology and the Centre for Integrative Physiology, Hugh Robson Building, University of EdinburghInstitute of Cell Biology and the Centre for Integrative Physiology, Hugh Robson Building, University of EdinburghMRC Centre for Reproductive Health, Queens Medical Research Institute, University of EdinburghInstitute of Cell Biology and the Centre for Integrative Physiology, Hugh Robson Building, University of EdinburghAbstract 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.https://doi.org/10.1038/s41598-018-25116-1 |
| spellingShingle | Yvonne L. Clarkson Marie McLaughlin Martin Waterfall Cheryl E. Dunlop Paul A. Skehel Richard A. Anderson Evelyn E. Telfer Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity Scientific Reports |
| title | Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity |
| title_full | Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity |
| title_fullStr | Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity |
| title_full_unstemmed | Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity |
| title_short | Initial characterisation of adult human ovarian cell populations isolated by DDX4 expression and aldehyde dehydrogenase activity |
| title_sort | initial characterisation of adult human ovarian cell populations isolated by ddx4 expression and aldehyde dehydrogenase activity |
| url | https://doi.org/10.1038/s41598-018-25116-1 |
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