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Myelodysplastic syndrome

Molecular and phenotypic characterization of CD133 and SSEA4 enriched very small embryonic-like stem cells in human cord blood

Leukemia volume 29, pages 1909–1917 (2015)Cite this article

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Abstract

Very small embryonic-like stem cells (VSELs) are immature primitive cells residing in adult and fetal tissues. This study describes enrichment strategy and molecular and phenotypic characterization of human cord blood VSELs. Flow cytometry analysis revealed that a majority of VSELs (LIN−/CD45−/CD34+) were present in the red blood cell (RBC) pellet after Ficoll-Hypaque centrifugation in contrast to the hematopoietic stem cells (LIN−/CD45+/CD34+) in the interphase layer. Thus, after lyses of RBCs, VSELs were enriched using CD133 and SSEA4 antibodies. These enriched cells were small in size (4–6 μm), spherical, exhibited telomerase activity and expressed pluripotent stem cell (OCT4A, OCT4, SSEA4, NANOG, SOX2, REX1), primordial germ cell (STELLA, FRAGILIS) as well as primitive hematopoietic (CD133, CD34) markers at protein and transcript levels. Heterogeneity was noted among VSELs based on subtle differences in expression of various markers studied. DNA analysis and cell cycle studies revealed that a majority of enriched VSELs were diploid, non-apoptotic and in G0/G1 phase, reflecting their quiescent state. VSELs also survived 5-fluorouracil treatment in vitro and treated cells entered into cell cycle. This study provides further support for the existence of pluripotent, diploid and relatively quiescent VSELs in cord blood and suggests further exploration of the subpopulations among them.

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Acknowledgements

We thank Dr Mukherjee, Gayatri Shinde, Sushma Khavale, Nivedita Dhavale, Dr Babu Rao, Dr Tamhankar, Harshvardhan Gawade and Dr Khatkhatay for their help. We also thank University Grants Commission, New Delhi, for support toward doctoral program of Ambreen Shaikh. Financial support for the study was provided by Indian Council of Medical Research, Government of India, New Delhi (Accession no. RA/214/01-2015).

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  1. Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India

    A Shaikh, P Nagvenkar, P Pethe & D Bhartiya

  2. Jaslok Hospital & Research Centre, Mumbai, India

    I Hinduja

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  1. A Shaikh
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Correspondence to D Bhartiya.

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Shaikh, A., Nagvenkar, P., Pethe, P. et al. Molecular and phenotypic characterization of CD133 and SSEA4 enriched very small embryonic-like stem cells in human cord blood. Leukemia 29, 1909–1917 (2015). https://doi.org/10.1038/leu.2015.100

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