α-Ketoglutarate Impairs the Long-Term Maintenance of Naïve Human Pluripotent Stem Cells
DING Liang1, XU Xueting2, QIN Baoming1,2*, WANG Lulu3*
This study aimed to investigate the decline in self-renewal capacity and pluripotency gene expression in nPSCs (naïve human pluripotent stem cells) during long-term passaging under 4CL conditions and to explore the underlying mechanisms. Additionally, the effect of αKG (alpha-ketoglutarate) on the steady-state maintenance of nPSCs was examined. Cell proliferation and pluripotency gene expression levels separately were assessed through cell growth curves and RT-qPCR during continuous passaging and after αKG treatment. Transcriptome sequencing was used to analyze differentially expressed genes after long-term passaging and αKG treatment, followed by functional analysis and motif prediction. Results indicated that cell proliferation and pluripotency gene expression gradually decreased during the continuous passaging of 4CL nPSCs, and were accompanied by changes in the expression of differentiation-related genes. Transcriptome analysis suggested that the decline in self-renewal might be associated with the activation of the p53 pathway, while the decrease in pluripotency gene expression could be linked to ETS and FOX family transcription factors. Short-term αKG treatment inhibited self-renewal without altering pluripotency gene expression, whereas long-term treatment improved self-renewal to some extent but further reduced pluripotency gene expression. In summary, the stability of 4CL nPSCs declines after long-term passaging, and this stability is further compromised by αKG treatment.
CN
EN