Abstract
Whether mitochondrial remodeling and metabolic reprogramming occur during the differentiation of human embryonic stem cells (hESCs) to definitive endoderm (DE) is unknown. We found that fragmented and punctate mitochondria in undifferentiated hESCs progressively fused into an extensive and branched network upon DE differentiation. Mitochondrial mass and mitochondrial DNA (mtDNA) content were significantly increased with the upregulated expression of mitochondrial biogenesis regulator PGC1-A upon DE differentiation, accompanied by the rise of the amount of ATP (2.5-fold) and its by-product reactive oxygen species (2.0-fold). We observed that in contrast to a shutoff of glycolysis, expressions of oxidative phosphorylation (OXPHOS) genes were increased, indicating that a transition from glycolysis to OXPHOS was tightly coupled to DE differentiation. In the meantime, we discovered that inhibition of TGF-β signaling led to impaired mitochondrial biogenesis and disturbed metabolic switch upon DE differentiation. Our work, for the first time, reports that TGF-β signaling–dependent mitochondrial biogenesis and metabolic reprogramming occur during early endodermal differentiation.
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Acknowledgments
We thank Duanqing Pei and Xiaodong Shu of the Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, for providing reagents or machines.
Funding
This work was supported by grants from the National Natural Science Foundation of China (grant number 31701183), Natural Science Foundation of Guangdong Province (grant number 2019A1515011324), and National Key R&D Program of China (grant number 2019YFA0111300).
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Editor: Tetsuji Okamoto
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Supplementary Figure 1
Short tandem repeat (STR) analysis of H1 cell line. (A) Match analysis. (B) Electropherogram of H1 cell line (PNG 942 kb)
Supplementary Data 1
Primers used for RT-qPCR and mtDNA copy number quantification. (XLSX 11 kb)
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Li, S., Huang, Q., Mao, J. et al. TGFβ-dependent mitochondrial biogenesis is activated during definitive endoderm differentiation. In Vitro Cell.Dev.Biol.-Animal 56, 378–385 (2020). https://doi.org/10.1007/s11626-020-00442-9
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DOI: https://doi.org/10.1007/s11626-020-00442-9