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Mouse Bone Marrow VSELs Exhibit Differentiation into Three Embryonic Germ Lineages and Germ & Hematopoietic Cells in Culture

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Abstract

Very small embryonic-like stem cells (VSELs) have been reported in various adult tissues, express pluripotent and primordial germ cells (PGCs) specific markers, are mobilized under stress/disease conditions, give rise to tissue committed progenitors and thus help regenerate and maintain homeostasis. The aim of the present study was to evaluate in vitro differentiation potential of VSELs using a quantitative approach. VSELs were collected from mouse bone marrow after 4 days of 5-fluorouracil (5-FU, 150 mg/Kg) treatment, further enriched by size based filtration and cultured on a feeder support in the presence of specific differentiation media. Cultured VSELs were found to differentiate into all three embryonic germ cell lineages, germ and hematopoietic cells after 14 days in culture. This was confirmed by studying Nestin, PDX-1, NKX2.5, DAZL, CD45 and other markers expression by various approaches. Very small, CD45 negative cells collected and enriched from GFP positive 5-FU treated mice bone marrow transitioned into CD45 positive cells in vitro thus demonstrating that VSELs can give rise to hematopoietic stem cells (HSCs). We envision that VSELs may be responsible for plasticity and ability of bone marrow cells to give rise to non-hematopoietic tissue progenitors of all 3 germ layers. Moreover the ability of VSELs to differentiate into germ cells as well as all the three lineages provides further evidence to support their pluripotent state and confirms developmental link between bone marrow VSELs and PGCs. The property of quiescence, no risk of teratoma formation and autologus source, make pluripotent VSELs a potential candidate to facilitate endogenous regeneration compared to cell replacement strategy envisioned using embryonic and induced pluripotent stem cells.

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Acknowledgements

We thank Gayatri Shinde and Sushma Khavale for their help in carrying out flow cytometry studies and to Shobha Sonawane and Reshma Gaonkar for their help with confocal studies. We acknowledge University Grants Commission (UGC), Government of India, New Delhi for support towards a doctoral program of AS. RG acknowledges Department of Science and Technology, Government of India under Woman Scientist Scheme-A [SR/WOSA/LS-1318] for fellowship.

NIRRH Accession No RA 373/05-2016.

Financial support for the study was provided by Indian Council of Medical Research, Government of India, New Delhi, INDIA.

Author Contributions

AS: Conception and design of the study, data collection, analysis and interpretation, manuscript writing. SA: Conception and design of the study, data collection, analysis and interpretation, manuscript writing. SK: Data collection, analysis, and interpretation, manuscript writing. DB: Conception and design of the study, data analysis and interpretation, manuscript writing. All authors read and approved the final version of the manuscript.

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  1. Stem Cell Biology Department, National Institute for Research in Reproductive Health (Indian Council of Medical Research), Jehangir Merwanji Street, Parel, Mumbai, 400 012, India

    Ambreen Shaikh, Sandhya Anand, Sona Kapoor, Ranita Ganguly & Deepa Bhartiya

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  1. Ambreen Shaikh
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  2. Sandhya Anand
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Correspondence to Deepa Bhartiya.

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Ambreen Shaikh, Sandhya Anand and Sona Kapoor equally contributed to the work

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Shaikh, A., Anand, S., Kapoor, S. et al. Mouse Bone Marrow VSELs Exhibit Differentiation into Three Embryonic Germ Lineages and Germ & Hematopoietic Cells in Culture. Stem Cell Rev and Rep 13, 202–216 (2017). https://doi.org/10.1007/s12015-016-9714-0

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