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Melatonin improves the reprogramming efficiency of murine‐induced pluripotent stem cells using a secondary inducible system
Author(s) -
Gao Shuai,
Wang ZhiLong,
Di KeQian,
Chang Gang,
Tao Li,
An Lei,
Wu FuJia,
Xu JiQiang,
Liu YingWei,
Wu ZhongHong,
Li XiangYun,
Gao Shaorong,
Tian JianHui
Publication year - 2013
Publication title -
journal of pineal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.881
H-Index - 131
eISSN - 1600-079X
pISSN - 0742-3098
DOI - 10.1111/jpi.12047
Subject(s) - melatonin , reprogramming , induced pluripotent stem cell , embryonic stem cell , biology , stem cell , homeobox protein nanog , andrology , microbiology and biotechnology , endocrinology , medicine , cell , genetics , gene
Abstract This study focused on the effect of melatonin on reprogramming with specific regard to the generation of induced pluripotent stem cells (i PSC s). 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 i PSC generation. Secondary fibroblasts collected from all‐i PSC 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 i PSC s induced by melatonin treatment ( M i PSC s) had the same characteristics as typical embryonic stem cells ( ESC s), including expression of the pluripotency markers O ct4, S ox2, and N anog, 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 MT 2 was detected in secondary fibroblasts, while M i PSC s and ESC s expressed MT 1 and MT 2 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 i PSC generation. These beneficial effects may be associated with inhibition of the p53‐mediated apoptotic pathway.