Embryonic Stem Cells Lacking DNA Methyltransferases Differentiate into Neural Stem Cells that Are Defective in Self-Renewal

Bong Jong Seo; Tae Kyung Hong; Sang Hoon Yoon; Jae Hoon Song; Sang Jun Uhm; Hyuk Song; Kwonho Hong; Hans Robert Schöler; Jeong Tae Do

doi:10.15283/ijsc22138

Embryonic Stem Cells Lacking DNA Methyltransferases Differentiate into Neural Stem Cells that Are Defective in Self-Renewal

Int J Stem Cells. 2023 Feb 28;16(1):44-51. doi: 10.15283/ijsc22138. Epub 2022 Oct 31.

Authors

Bong Jong Seo¹, Tae Kyung Hong^{1

2}, Sang Hoon Yoon^{1

2}, Jae Hoon Song¹, Sang Jun Uhm³, Hyuk Song¹, Kwonho Hong¹, Hans Robert Schöler⁴, Jeong Tae Do^{1

2}

Affiliations

¹ Department of Stem Cell and Regenerative Biotechnology, Konkuk Institute of Technology, Konkuk University, Seoul, Korea.
² 3D Tissue Culture Research Center, Konkuk University, Seoul, Korea.
³ Department of Animal Science, Sangji University, Wonju, Korea.
⁴ Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany.

Abstract

Background and objectives: DNA methyltransferases (Dnmts) play an important role in regulating DNA methylation during early developmental processes and cellular differentiation. In this study, we aimed to investigate the role of Dnmts in neural differentiation of embryonic stem cells (ESCs) and in maintenance of the resulting neural stem cells (NSCs).

Methods and results: We used three types of Dnmt knockout (KO) ESCs, including Dnmt1 KO, Dnmt3a/3b double KO (Dnmt3 DKO), and Dnmt1/3a/3b triple KO (Dnmt TKO), to investigate the role of Dnmts in neural differentiation of ESCs. All three types of Dnmt KO ESCs could form neural rosette and differentiate into NSCs in vitro. Interestingly, however, after passage three, Dnmt KO ESC-derived NSCs could not maintain their self-renewal and differentiated into neurons and glial cells.

Conclusions: Taken together, the data suggested that, although deficiency of Dnmts had no effect on the differentiation of ESCs into NSCs, the latter had defective maintenance, thereby indicating that Dnmts are crucial for self-renewal of NSCs.

Keywords: DNA methyltransferases; Neural stem cells; Neuronal and glial differentiation.

Grants and funding

Acknowledgments This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) of the Republic of Korea (grant no. 2020R1A2C3007562) and Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET), funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (grant no. 322006-05-01-CG000).