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      Single-cell analysis of human ovarian cortex identifies distinct cell populations but no oogonial stem cells

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          Abstract

          The human ovary orchestrates sex hormone production and undergoes monthly structural changes to release mature oocytes. The outer lining of the ovary (cortex) has a key role in defining fertility in women as it harbors the ovarian reserve. It has been postulated that putative oogonial stem cells exist in the ovarian cortex and that these can be captured by DDX4 antibody isolation. Here, we report single-cell transcriptomes and cell surface antigen profiles of over 24,000 cells from high quality ovarian cortex samples from 21 patients. Our data identify transcriptional profiles of six main cell types; oocytes, granulosa cells, immune cells, endothelial cells, perivascular cells, and stromal cells. Cells captured by DDX4 antibody are perivascular cells, not oogonial stem cells. Our data do not support the existence of germline stem cells in adult human ovaries, thereby reinforcing the dogma of a limited ovarian reserve.

          Abstract

          The outer lining or cortex of the human ovary determines fertility and has been postulated to contain oogonial stem cells. Here, the authors generate a single-cell map of the adult human ovarian cortex and show that DDX4 labels perivascular cells but no oogonial stem cells.

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          Most cited references47

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          A practical guide to single-cell RNA-sequencing for biomedical research and clinical applications

          RNA sequencing (RNA-seq) is a genomic approach for the detection and quantitative analysis of messenger RNA molecules in a biological sample and is useful for studying cellular responses. RNA-seq has fueled much discovery and innovation in medicine over recent years. For practical reasons, the technique is usually conducted on samples comprising thousands to millions of cells. However, this has hindered direct assessment of the fundamental unit of biology—the cell. Since the first single-cell RNA-sequencing (scRNA-seq) study was published in 2009, many more have been conducted, mostly by specialist laboratories with unique skills in wet-lab single-cell genomics, bioinformatics, and computation. However, with the increasing commercial availability of scRNA-seq platforms, and the rapid ongoing maturation of bioinformatics approaches, a point has been reached where any biomedical researcher or clinician can use scRNA-seq to make exciting discoveries. In this review, we present a practical guide to help researchers design their first scRNA-seq studies, including introductory information on experimental hardware, protocol choice, quality control, data analysis and biological interpretation.
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            Single-Cell RNA-Seq Analysis Maps Development of Human Germline Cells and Gonadal Niche Interactions.

            Human fetal germ cells (FGCs) are precursors to sperm and eggs and are crucial for maintenance of the species. However, the developmental trajectories and heterogeneity of human FGCs remain largely unknown. Here we performed single-cell RNA-seq analysis of over 2,000 FGCs and their gonadal niche cells in female and male human embryos spanning several developmental stages. We found that female FGCs undergo four distinct sequential phases characterized by mitosis, retinoic acid signaling, meiotic prophase, and oogenesis. Male FGCs develop through stages of migration, mitosis, and cell-cycle arrest. Individual embryos of both sexes simultaneously contain several subpopulations, highlighting the asynchronous and heterogeneous nature of FGC development. Moreover, we observed reciprocal signaling interactions between FGCs and their gonadal niche cells, including activation of the bone morphogenic protein (BMP) and Notch signaling pathways. Our work provides key insights into the crucial features of human FGCs during their highly ordered mitotic, meiotic, and gametogenetic processes in vivo.
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              Intraovarian Control of Early Folliculogenesis

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                Author and article information

                Contributors
                Fredrik.Lanner@ki.se
                Pauliina.Damdimopoulou@ki.se
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                2 March 2020
                2 March 2020
                2020
                : 11
                : 1147
                Affiliations
                [1 ]ISNI 0000 0000 9241 5705, GRID grid.24381.3c, Division of Obstetrics and Gynecology, Department of Clinical Science, Intervention and Technology, , Karolinska Institutet and Karolinska University Hospital, ; Stockholm, Sweden
                [2 ]ISNI 0000 0004 1937 0626, GRID grid.4714.6, Department of Biosciences and Nutrition, , Karolinska Institutet, ; Stockholm, Sweden
                [3 ]ISNI 0000 0004 1937 0626, GRID grid.4714.6, Center of Hematology and Regenerative Medicine, Department of Medicine, , Karolinska Institutet, ; Stockholm, Sweden
                [4 ]ISNI 0000 0004 1937 0626, GRID grid.4714.6, Bioinformatics and Expression Analysis core facility, Karolinska Institutet, ; Stockholm, Sweden
                [5 ]ISNI 0000 0001 2292 3357, GRID grid.14848.31, Department of Medicine, Centre de recherche du CHUM, , University of Montreal, ; Montreal, Canada
                [6 ]GRID grid.440124.7, Department of Gynecology and Obstetrics, , Visby hospital, ; Visby, Sweden
                [7 ]ISNI 0000 0004 0410 2071, GRID grid.7737.4, Folkhälsan Research Institute, Helsinki, and Stem Cells and Metabolism Research Program, , University of Helsinki, ; Helsinki, Finland
                [8 ]ISNI 0000 0004 1937 0626, GRID grid.4714.6, Ming Wai Lau Center for Reparative Medicine, Stockholm node, Karolinska Institutet, ; Stockholm, Sweden
                [9 ]ISNI 0000 0001 2164 9667, GRID grid.419681.3, Present Address: Flow Cytometry Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, ; Bethesda, MD USA
                Author information
                http://orcid.org/0000-0002-6266-1248
                http://orcid.org/0000-0002-8915-9282
                http://orcid.org/0000-0002-3221-8667
                http://orcid.org/0000-0001-5395-3807
                http://orcid.org/0000-0003-1974-0271
                http://orcid.org/0000-0002-2771-7445
                http://orcid.org/0000-0001-8458-0855
                Article
                14936
                10.1038/s41467-020-14936-3
                7052271
                32123174
                7059b97f-fba0-461b-8675-1e59fff7d49d
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 22 October 2019
                : 10 February 2020
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100004359, Vetenskapsrådet (Swedish Research Council);
                Funded by: FundRef https://doi.org/10.13039/501100007263, Astellas Foundation for Research on Metabolic Disorders;
                Funded by: FundRef https://doi.org/10.13039/501100004063, Knut och Alice Wallenbergs Stiftelse (Knut and Alice Wallenberg Foundation);
                Funded by: FundRef https://doi.org/10.13039/501100006313, Barncancerfonden (Swedish Childhood Cancer Foundation);
                Funded by: FundRef https://doi.org/10.13039/100007459, Ragnar Söderbergs stiftelse (Ragnar Söderberg Foundation);
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                Uncategorized
                cell biology,stem-cell niche
                Uncategorized
                cell biology, stem-cell niche

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