Antioxidant Supplementation Reduces Genomic Aberrations in Human Induced Pluripotent Stem Cells

Abstract

SummarySomatic cells can be reprogrammed to induced pluripotent stem cells (iPSCs) using oncogenic transcription factors. However, this method leads to genetic aberrations in iPSCs via unknown mechanisms, which may limit their clinical use. Here, we demonstrate that the supplementation of growth media with antioxidants reduces the genome instability of cells transduced with the reprogramming factors. Antioxidant supplementation did not affect transgene expression level or silencing kinetics. Importantly, iPSCs made with antioxidants had significantly fewer de novo copy number variations, but not fewer coding point mutations, than iPSCs made without antioxidants. Our results suggest that the quality and safety of human iPSCs might be enhanced by using antioxidants in the growth media during the generation and maintenance of iPSCs.

Stem cells and the impact of ROS signaling

“…Moreover, new methods of small molecule-mediated iPSC generation have been shown to modulate the transition to aerobic glycolysis (Zhu et al, 2010). Although the precise effect of ROS on signaling pathways during the reprogramming process has not been evaluated, levels of ROS appear to increase during reprogramming and to cause damage to DNA, which can be minimized by the addition of NAC (Ji et al, 2014). The efficiency of reprogramming and continued maintenance of iPSCs can also be improved under low O 2 conditions (Ezashi et al, 2005).…”

Inducing pluripotency in vitro: recent advances and highlights in induced pluripotent stem cells generation and pluripotency reprogramming

“…Some of the recent studies have sought to address nutritional requirements for pluripotency reprogramming and determined that some common dietary supplements have noteworthy potential for improving reprogramming efficiency and iPSC quality (Table , Fig. ).…”

“…Relatively higher numbers of authentic-iPSCs following Vc treatment may in part be due to Vc-mediated transformation of partially reprogrammed pre-iPSCs into fully reprogrammed iPSCs (111). Furthermore, Stadtfeld et al 113,114 demonstrated that Vc treatment substantially improved the quality of iPSCs, reviving full developmental potential and unaltered genomic status (112). Generally, OSKM-transduced miPSCs bear altered epigenetic signatures at the imprinted Dlk1-Dio3 gene locus which modulates their ability to generate iPSC-derived adult mice through tetraploid complementation.…”

“…However, supplementing reprogramming media with Vc restores H3 lysine 4 trimethylation at Dlk1-Dio3 loci of the OSKM-iPSCs, reviving full pluripotency (112). Also, supplementing reprogramming media with antioxidants, such as N-acetyl-cysteine (NAC), can substantially reduce genomic instability of iPSCs, one serious side effect of pluripotency reprogramming (113,114). NAC tends to inhibit rapid build-up of reactive oxygen species and reduces de novo copy number variations in OSKM-iPSCs, due to metabolic shift during early phases of the reprogramming (113).…”

Concise Review: Genomic Instability in Human Stem Cells: Current Status and Future Challenges

“…In line with these findings, hiPSCs also seem to form teratomas faster and with higher aggressiveness than hESCs, irrespective of the site of administration or engraftment . Two new studies have observed that mitigation of oxidative stress during reprogramming and early passaging by using antioxidant supplements in the growth media was able to reduce genome instability in hiPSCs .…”