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Stem Cells in Adult Mice Ovaries Form Germ Cell Nests, Undergo Meiosis, Neo-oogenesis and Follicle Assembly on Regular Basis During Estrus Cycle

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A Correction to this article was published on 19 October 2021

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Abstract

Very small embryonic-like (VSELs) and ovarian (OSCs) stem cells are located in adult mammalian ovary surface epithelium (OSE). OSCs can expand long-term and differentiate into oocyte-like structures in vitro and have resulted in birth of fertile pups. Lineage tracing studies have provided evidence to suggest OSCs differentiation into oocytes in vivo. But how these stem cells function under normal physiological conditions has not yet been well worked out. Besides studying STRA-8 and SCP-3 expression in enzymatically isolated OSE cells smears, mice were injected BrdU to track mitosis, meiosis and follicle assembly. H&E stained OSE cells during late diestrus and proestrus showed VSELs undergoing asymmetrical cell divisions to give rise to slightly bigger OSCs which in turn underwent symmetrical cell divisions followed by clonal expansion (rapid expansion with incomplete cytokinesis) during early estrus to form germ cell nests (GCN). OCT-4, SSEA-1, MVH and DAZL positive cells in GCN expressed Erα, Erβ and FSHR, were interconnected by ring canals (TEX-14), showed mitochondrial aggregation (Cytochrome C) and Balbiani Body (TRAL). Apoptosis in ‘nurse’ cells was marked by PARP and putative oocytes were clearly visualized. BrdU was detected in cells undergoing mitosis/meiosis and also in an oocyte of secondary follicle. FACS sorted, green fluorescent protein (GFP) positive VSELs upon transplantation resulted in GFP positive GCN suggesting crucial role for VSELs in adult ovaries. Results suggest that various events described during oogenesis and follicle assembly in fetal ovaries are recapitulated on regular basis in adult ovary and result in the formation of follicles.

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Acknowledgements

We acknowledge the help provided by Dr SM Metkari at the Animal Experimental Facility, NIRRH for transplanting VSELs in mice ovaries. Help from Gayatri and Sushma for flow cytometry and Shobha, Reshma and Swati for confocal microscopy is acknowledged.

Funding

Financial support was provided by Indian Council of Medical Research, Government of India, New Delhi, India. DS acknowledges SRF fellowship from ICMR (#10832).

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Authors and Affiliations

  1. Stem Cell Biology Department, ICMR- National Institute for Research in Reproductive Health, Jehangir Merwanji Street, Parel, Mumbai, 400 012, India

    Diksha Sharma & Deepa Bhartiya

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  1. Diksha Sharma
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  2. Deepa Bhartiya
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DS performed the experiments, generated results, data interpretation and manuscript preparation. DB designed the study, arranged funds, data interpretation and manuscript preparation. Both authors agree to the final version of submitted manuscript.

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Correspondence to Deepa Bhartiya.

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All experiments carried out in the present study were approved by NIRRH Institutional Animal Ethics Committee (78/GO/ReBi/SL/99/CPCSEA dated-11/03/1999).

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Institute accession number NIRRH No. REV/1074/05-2021.

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Authors declare no conflict of interest.

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The original online version of this article was revised: The wrong Supplementary file for Supplementary file 1 was originally published with this article; it has now been replaced with the correct file.

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Sharma, D., Bhartiya, D. Stem Cells in Adult Mice Ovaries Form Germ Cell Nests, Undergo Meiosis, Neo-oogenesis and Follicle Assembly on Regular Basis During Estrus Cycle. Stem Cell Rev and Rep 17, 1695–1711 (2021). https://doi.org/10.1007/s12015-021-10237-4

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