title: Melatonin improves the reprogramming efficiency of murine-induced pluripotent stem cells using a secondary inducible system.
abstract: This study focused on the effect of melatonin on reprogramming with specific regard to the generation of induced pluripotent stem cells (iPSCs). Here, a secondary inducible system, which is more accurate and suitable for studying the involvement of chemicals in reprogramming efficiency, was used to evaluate the effect of melatonin on mouse iPSC generation. Secondary fibroblasts collected from all-iPSC mice through tetraploid complementation were cultured in induction medium supplemented with melatonin at different concentrations (0, 10(-6), 10(-7), 10(-8), 10(-9), or 10(-10 )m) or with vitamin C (50 μg/mL) as a positive control. Compared with untreated group (0.22 ± 0.04% efficiency), 10(-8) (0.81 ± 0.04%), and 10(-9 )m (0.83 ± 0.08%) melatonin supplementation significantly improved reprogramming efficiency (P < 0.05). Moreover, we verified that the iPSCs induced by melatonin treatment (MiPSCs) had the same characteristics as typical embryonic stem cells (ESCs), including expression of the pluripotency markers Oct4, Sox2, and Nanog, the ability to form teratomas and all three germ layers of the embryo, as well as produce chimeric mice with contribution to the germ line. Interestingly, only the melatonin receptor MT2 was detected in secondary fibroblasts, while MiPSCs and ESCs expressed MT1 and MT2 receptors. Furthermore, during the early stage of reprogramming, expression of the apoptosis-related genes p53 and p21 was lower in the group treated with 10(-9) m melatonin compared with the untreated controls. In conclusion, melatonin supplementation enhances the efficiency of murine iPSC generation. These beneficial effects may be associated with inhibition of the p53-mediated apoptotic pathway.
authors.affiliation: Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture, National Engineering Laboratory for Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China; National Institute of Biological Sciences, NIBS, Beijing, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Gao
authors.forename: Zhi-Long
authors.initials: ZL
authors.lastname: Wang
authors.forename: Ke-Qian
authors.initials: KQ
authors.lastname: Di
authors.forename: Gang
authors.initials: G
authors.lastname: Chang
authors.forename: Li
authors.initials: L
authors.lastname: Tao
authors.forename: Lei
authors.initials: L
authors.lastname: An
authors.forename: Fu-Jia
authors.initials: FJ
authors.lastname: Wu
authors.forename: Ji-Qiang
authors.initials: JQ
authors.lastname: Xu
authors.forename: Ying-Wei
authors.initials: YW
authors.lastname: Liu
authors.forename: Zhong-Hong
authors.initials: ZH
authors.lastname: Wu
authors.forename: Xiang-Yun
authors.initials: XY
authors.lastname: Li
authors.forename: Shaorong
authors.initials: S
authors.lastname: Gao
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Tian
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002478:Cells, Cultured; D002678:Chimera / Q000235:genetics / Q000378:metabolism; D005260:Female; D005347:Fibroblasts; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D008297:Male; D008550:Melatonin / Q000494:pharmacology*; D051379:Mice; D008813:Mice, Inbred ICR; D016513:Mice, SCID; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics / Q000378:metabolism; D044094:Receptors, Melatonin / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D050814:Octamer Transcription Factor-3; D044094:Receptors, Melatonin; D016159:Tumor Suppressor Protein p53; D016283:Proto-Oncogene Proteins p21(ras); D008550:Melatonin
keywords: apoptosis; induced pluripotent stem cells; melatonin; membrane receptor; secondary inducible system