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Define of Optimal Addition Period of Osteogenic Peptide to Accelerate the Osteogenic Differentiation of Human Pluripotent Stem Cells

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period.

Methods:

Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7.

Results:

Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs.

Conclusion:

The differentiation efficiency of cells could be improved by determining the optimal treatment period. Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.

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Data availability

The data presented in this study are available on request from all the authors.

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Acknowledgements

This work was funded by the Foundation for the Talents of Innovative and Entrepreneurial of Lanzhou (NO. 2021-RC-127), the Open Subject Foundation of Key Laboratory of Dental Maxillofacial Reconstruction and Biological Intelligence Manufacturing, Fundamental Research Funds for the Central Universities (NO. lzujbky-2021-kb05 and lzujbky-2021-ey14), the Lanzhou University Hospital of Stomatology Research Support Fund, the Natural Science Foundation of Gansu Province (NO. 21JR7RA863, NO. 20JR5RA150 and 22JR5RA499), and the China Postdoctoral Science Foundation (NO.2022M721443).

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Author notes

  1. Yameng Song and Hongjiao Li have contributed equally.

Authors and Affiliations

  1. School and Hospital of Stomatology, Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Yameng Song, Hongjiao Li, Zixuan Wang, Jing Li, Lu Wang, Lingzi Liao, Shengqin Ma, Bin Liu & Ping Zhou

  2. School of Life Sciences, Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Jiamin Shi

  3. Lanzhou Hospital of Stomatology, Lanzhou, 730000, People’s Republic of China

    Yun Zhang & Yaling Yang

  4. Department of Orthopedics, Lanzhou University Second Hospital, No.82 Cuiyingmen Street, Lanzhou, 730030, Gansu, People’s Republic of China

    Ping Zhou

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Contributions

PZ, YLY and BL contributed to designing the study and critically revised the manuscript. PZ, YMS, HJL, LJ and JMS contributed to drafting and reviewing the manuscript. YMS, LW, ZXW, LZL, SQM, SZL and YZ performed all the experimental work. YMS contributed to performing the statistical analysis and interpreting the results. All authors have read and approved the article and have due diligence to ensure the integrity of the manuscript. Neither the entire manuscript nor any part of its content has been published or accepted elsewhere.

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Correspondence to Bin Liu, Yaling Yang or Ping Zhou.

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Song, Y., Li, H., Wang, Z. et al. Define of Optimal Addition Period of Osteogenic Peptide to Accelerate the Osteogenic Differentiation of Human Pluripotent Stem Cells. Tissue Eng Regen Med 21, 291–308 (2024). https://doi.org/10.1007/s13770-023-00597-y

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