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Osteogenic Differentiation of Menstrual Blood-Derived Stem Cells by Optogenetics

Abstract

Purpose

Menstrual blood-derived stem cells (MenSCs), a novel source of mesenchymal stem cells (MSC)s which can be extracted from the menstrual fluid by a simple and non-invasive approach, and without ethical concerns. Optogenetics is a biological tool for controlling ion flux by light in various cells, including stem cells. This study aims to evaluate the osteogenic differentiation of MenSCs by optogenetic manipulation.

Methods

The MenSCs were transduced with lentivirus vector carrying channelrhodopsin-2 [hChR2 (H134R)]. ChR2-expressing MenSCs were illuminated with blue light (470 nm) with 1 mW power for 1 h. Several control groups, including non-ChR2-expressing MenSCs in conventional culture medium and osteogenic medium with dark and optical control, were subjected in this study. A day after light illumination, the viability of cells and intracellular calcium (Ca2+) level was evaluated by the MTT assay, and spectrophotometer, respectively. The Ca2+ deposition and alkaline phosphatase activity also were examined on days 7, 14, and 21. The expression of osteogenic genes Runx2, osteopontin, and osteocalcin were analyzed by quantitative PCR.

Results

72 h after infection, MenSCs were effectively transduced using lentiviral vector. In comparison to other groups, optogenetic activated ChR2-expressing MenSCs showed significantly higher viability in the MTT experiment. Additionally, optogenetic stimulation increased intracellular Ca2+ concentration and deposition, as well as alkaline phosphatase activity (ALP). Optical stimulation also promoted the production of osteogenic genes such Runx2, osteocalcin, and osteopontin.

Conclusion

Optical stimulation could increase the intracellular Ca2+ level, ALP activity, cell viability, and up-regulation of Runx2, osteocalcin, and osteopontin, which play an important role during osteogenesis. Hence, the results of present study may provide a new strategy forbone regeneration and regenerative medicine.

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Acknowledgements

This project was funded by a grant from National Institute for Medical Research Development (NIMAD), Tehran, Iran (grant No.988136).

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Correspondence to Gholamreza Esmaeeli Djavid.

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Neghab, H.K., Djavid, G.E., Azadeh, S.S. et al. Osteogenic Differentiation of Menstrual Blood-Derived Stem Cells by Optogenetics. J. Med. Biol. Eng. (2022). https://doi.org/10.1007/s40846-022-00714-7

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  • DOI: https://doi.org/10.1007/s40846-022-00714-7

Keywords

  • Optogenetics
  • Osteogenic differentiation
  • Menstrual blood-derived stem cells
  • Stem cells
  • Channelrhodopsin