title: Ethanol Inactivated Mouse Embryonic Fibroblasts Maintain the Self-Renew and Proliferation of Human Embryonic Stem Cells.
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.
authors.affiliation: 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.
authors.forename: Boxian
authors.initials: B
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Center of Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
authors.forename: Song
authors.initials: S
authors.lastname: Ning
authors.affiliation: Department of Pediatrics, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu Province, 210029, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Zhuang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Center of Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Center of Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
authors.forename: Yugui
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Department of Human Genetics, David Geffen School of Medicine, UCLA, Los Angeles, California, 90095, United States of America.
authors.forename: Guoping
authors.initials: G
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Reproductive Medicine, Center of Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
authors.forename: Lianju
authors.initials: L
authors.lastname: Qin
authors.affiliation: 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.
authors.forename: Jiayin
authors.initials: J
authors.lastname: Liu
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D000066673:Cell Self Renewal; D018920:Coculture Techniques; D015925:Cryopreservation; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D000066449:Human Embryonic Stem Cells / Q000502:physiology*; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D039904:Pluripotent Stem Cells / Q000502:physiology
chemical_list: D015415:Biomarkers; D000431:Ethanol