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In vitro differentiation of primed human induced pluripotent stem cells into primordial germ cell-like cells

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Abstract

Background

Previous studies have shown significant results in the differentiation of mouse-induced pluripotent stem cells (miPSCs) into primordial germ cell-like cells (PGCLCs) and that human iPSCs (hiPSCs) can also differentiate into PGCLCs; however, the efficiency of PGCLC induction from hiPSCs is < 5%. In this study, we examined a new protocol to differentiate hiPSCs into PGCLCs.

Methods and Results

hiPSCs-derived embryoid bodies (EBs) were exposed to differentiate inducing factors, bone morphogenetic protein 4 (BMP4), and retinoic acid (RA) for 6 days. Cell differentiation was assessed by reverse transcriptase–polymerase chain reaction (RT-PCR) and immunofluorescence (IF) studies. Our results showed increased expression of the PRDM1 gene on the first day of differentiation. On other days, DAZL, VASA, and STRA8 genes increased, and the expression of PRDM1, NANOG, and OCT4 genes decreased. The expression of VASA, C-KIT, and STRA8 proteins was confirmed by IF. A flow cytometry analysis revealed that ~ 60% of differentiated cells were VASA- and STRA8-positive.

Conclusion

EB formation and constant exposure of EBs to BMP4 and RA lead to the differentiation of hiPSCs into PGCLCs.

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Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

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Authors and Affiliations

  1. Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Jalal Ale Ahmad Highway, P.O. Box: 14115-111, Tehran, Iran

    Mehrafarin Kiani, Mansoureh Movahedin & Iman Halvaei

  2. Department of Hematology and Stem Cell Therapy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Masoud Soleimani

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  1. Mehrafarin Kiani
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  2. Mansoureh Movahedin
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  3. Iman Halvaei
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  4. Masoud Soleimani
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Contributions

MK performed all the experiments, data analysis and drafting the manuscript. MM was supervised the project, editing the manuscript and participated in the finalization of the manuscript. MS prepared the iPSCs. IH participated in statistical analysis and editing the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Mansoureh Movahedin.

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Kiani, M., Movahedin, M., Halvaei, I. et al. In vitro differentiation of primed human induced pluripotent stem cells into primordial germ cell-like cells. Mol Biol Rep 50, 1971–1979 (2023). https://doi.org/10.1007/s11033-022-08012-w

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