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.

Melatonin promotes development of haploid germ cells from early developing spermatogenic cells of Suffolk sheep under in vitro condition

“…Melatonin has been known as a pleiotropic molecule. It is a potent free radical scavenger and antioxidant , and it also regulates cell proliferation and differentiation . Both melatonin membrane receptors 1 (MT1) and 2 (MT2) are G protein‐coupled receptors and regulate the activation of adenylate cyclase (AC) and phospholipase C (PLC) .…”

Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

“…Studies have shown evidences that melatonin can enhance the production of iPSCs due to its antioxidant activity [8, 9]. In human adult MSCs, for instance, melatonin combined with the extracellular matrix can enhance cellular alkaline phosphatase (AP) activity, induce osteogenesis via the melatonin receptor 2 (MT2), and also reverse stress-induced MSC injuries [10–13].…”

Embryonic stem‐like cells from rabbit blastocysts cultured with melatonin could differentiate into three germ layers in vitro and in vivo

“…Recent reports suggest that melatonin supplementation enhances the quality of porcine embryos (Do et al, ) and bovine cloned embryos (Su et al, ). Melatonin supplementation also significantly improved the reprogramming efficiency of murine induced pluripotent cells (Gao et al, ). We therefore, asked if melatonin could enhance the derivation and maintenance of ESCs from rabbits using established methods.…”