Ethanol Inactivated Mouse Embryonic Fibroblasts Maintain the Self-Renew and Proliferation of Human Embryonic Stem Cells

Boxian Huang; Song Ning; Lili Zhuang; Chunyan Jiang; Yugui Cui; Guoping Fan; Lianju Qin; Jiayin Liu

doi:10.1371/journal.pone.0130332

Ethanol Inactivated Mouse Embryonic Fibroblasts Maintain the Self-Renew and Proliferation of Human Embryonic Stem Cells

PLoS One. 2015 Jun 19;10(6):e0130332. doi: 10.1371/journal.pone.0130332. eCollection 2015.

Authors

Boxian Huang¹, Song Ning², Lili Zhuang³, Chunyan Jiang², Yugui Cui², Guoping Fan⁴, Lianju Qin², Jiayin Liu¹

Affiliations

¹ School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210038, China; State Key Laboratory of Reproductive Medicine, Center of Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
² State Key Laboratory of Reproductive Medicine, Center of Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
³ Department of Pediatrics, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu Province, 210029, China.
⁴ Department of Human Genetics, David Geffen School of Medicine, UCLA, Los Angeles, California, 90095, United States of America.

Abstract

Conventionally, mouse embryonic fibroblasts (MEFs) inactivated by mitomycin C or irradiation were applied to support the self-renew and proliferation of human embryonic stem cells (hESCs). To avoid the disadvangtages of mitomycin C and irradiation, here MEFs were treated by ethanol (ET). Our data showed that 10% ET-inactivated MEFs (eiMEFs) could well maintain the self-renew and proliferation of hESCs. hESCs grown on eiMEFs expressed stem cell markers of NANOG, octamer-binding protein 4 (OCT4), stage-specific embryonic antigen-4 (SSEA4) and tumour related antigen-1-81 (TRA-1-81), meanwhile maintained normal karyotype after long time culture. Also, hESCs cocultured with eiMEFs were able to form embryoid body (EB) in vitro and develop teratoma in vivo. Moreover, eiMEFs could keep their nutrient functions after long time cryopreservation. Our results indicate that the application of eiMEF in hESCs culture is safe, economical and convenient, thus is a better choice.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biomarkers / metabolism
Cell Culture Techniques
Cell Proliferation
Cell Self Renewal*
Coculture Techniques
Cryopreservation
Ethanol / pharmacology*
Female
Fibroblasts / drug effects
Fibroblasts / physiology*
Human Embryonic Stem Cells / physiology*
Humans
Mice
Mice, Inbred ICR
Pluripotent Stem Cells / physiology

Substances

Biomarkers
Ethanol

Associated data

figshare/10.6084/M9.FIGSHARE.1431806
figshare/10.6084/M9.FIGSHARE.1431818
figshare/10.6084/M9.FIGSHARE.14318187

Grants and funding

This work was supported by the grants from the State Major Research Program of China (2012CB944902), National Natural Science Foundation of China (81370764) Jiangsu projects (BL2012009, FXK201221 and the PAPD project) and China Scholarship Council (201307060009).