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Osteogenic differentiation of human induced pluripotent stem cell in the presence of testosterone and 17 β-estradiol in vitro

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Abstract

Recently, numerous scientific approaches have been explored to treat various diseases using stem cells. In 2006, induced pluripotent stem cell (iPSC) were introduced by Takahashi and Yamanaka and showed the potential of self-renewing and differentiation into all types of targeted cells in vitro. In this investigation, we studied the effect of testosterone (T) individually or in the presence of 17 β-estradiol (E2) on osteogenic differentiation of human iPSC (hiPSC) during 2 wk. The optimal concentrations of sex steroid hormones were examined by MTT assay and acridine orange (AO) staining. The impact of E2 and T either individually or together as a combination was examined by ALP activity; the content of total mineral calcium, by von Kossa and alizarin red staining. Additionally, the expression rate of osteogenic specific markers was studied via real-time RT-PCR and immunocytochemistry analyses at day 14 of differentiation. The obtained results illustrated that the differentiation medium supplemented with T-E2 increased not only the ALP enzyme activity and the content of calcium but also the osteogenic-related gene and protein expressions on the 14th day. Furthermore, the results were confirmed by mineralized matrix staining. In conclusion, these data suggest that T could be used as an effective factor for osteogenic induction of hiPSCs combined with the E2 in bone regeneration.

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Abbreviations

AR:

Androgen receptor

AO:

Acridine orange

AA:

Amino acids

cAMP:

Cyclic adenosine monophosphate

EB:

Ethidium bromide

ESCs:

Embryonic stem cells

E2:

17 β-Estradiol

hiPSCs:

Human induced pluripotent stem cells

T:

Testosterone

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Acknowledgements

We thank Ms. Elahe Eftekhari for her assistance with the preparation of this manuscript.

Author information

Authors and Affiliations

  1. Stem Cell and Regenerative Medicine Center of Excellence, Tehran University of Medical Sciences, Tehran, Iran

    Reyhaneh Yarmohammadi & Mohammad Amir Amirkhani

  2. Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, P. O. Box, 15719-14911, Tehran, Iran

    Marzieh Ghollasi & Maryam Soltanyzadeh

  3. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

    Fatemeh Kheirollahzadeh

  4. Department of Cellular and Molecular Biology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

    Masoumeh Heshmati

Authors
  1. Reyhaneh Yarmohammadi
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  2. Marzieh Ghollasi
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  3. Fatemeh Kheirollahzadeh
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  4. Maryam Soltanyzadeh
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  5. Masoumeh Heshmati
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  6. Mohammad Amir Amirkhani
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Correspondence to Marzieh Ghollasi.

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The authors declare no competing interests.

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This article does not contain any studies with human participants or animals performed by any of the authors. The hiPSCs were purchased from The Stem Cell Technology Research Center.

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Yarmohammadi, R., Ghollasi, M., Kheirollahzadeh, F. et al. Osteogenic differentiation of human induced pluripotent stem cell in the presence of testosterone and 17 β-estradiol in vitro. In Vitro Cell.Dev.Biol.-Animal 58, 179–188 (2022). https://doi.org/10.1007/s11626-022-00652-3

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