2. Academic Validation
  3. Mitochondrial homeostasis regulates definitive endoderm differentiation of human pluripotent stem cells

Mitochondrial homeostasis regulates definitive endoderm differentiation of human pluripotent stem cells

  • Cell Death Discov. 2022 Feb 17;8(1):69. doi: 10.1038/s41420-022-00867-z.
Jing Lv  # 1,  Ying Yi  # 1,  Yan Qi 2,  Chenchao Yan 1,  Wenwen Jin 1,  Liming Meng 1,  Donghui Zhang 3,  Wei Jiang 4 5 6
Affiliations

Affiliations

  • 1 Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, 430071, China.
  • 2 State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Science, Hubei University, Wuhan, 430062, China.
  • 3 State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Science, Hubei University, Wuhan, 430062, China. [email protected].
  • 4 Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, 430071, China. [email protected].
  • 5 Human Genetics Resource Preservation Center of Wuhan University, Wuhan, 430071, China. [email protected].
  • 6 Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, 430071, China. [email protected].
  • # Contributed equally.
PMID: 35177589 DOI: 10.1038/s41420-022-00867-z
Abstract

Cellular organelles play fundamental roles in almost all cell behaviors. Mitochondria have been reported to be functionally linked to various biological processes, including reprogramming and pluripotency maintenance. However, very little about the role of mitochondria has been revealed in human early development and lineage specification. Here, we reported the characteristics and function of mitochondria during human definitive endoderm differentiation. Using a well-established differentiation system, we first investigated the change of mitochondrial morphology by comparing undifferentiated pluripotent stem cells, the intermediate mesendoderm cells, and differentiated endoderm cells, and found that mitochondria were gradually elongated and matured along differentiation. We further analyzed the expression pattern of mitochondria-related genes by RNA-seq, indicating that mitochondria became active during differentiation. Supporting this notion, the production of adenosine triphosphate (ATP) and Reactive Oxygen Species (ROS) was increased as well. Functionally, we utilized chemicals and genome editing techniques, which could interfere with mitochondrial homeostasis, to determine the role of mitochondria in human endoderm differentiation. Treatment with mitochondrial inhibitors, or genetic depletion of mitochondrial transcription factor A (TFAM), significantly reduced the differentiation efficiency of definitive endoderm. In addition, the defect in endoderm differentiation due to dysfunctional mitochondria could be restored to some extent by the addition of ATP. Moreover, the clearance of excessive ROS due to dysfunctional mitochondria by N-acetylcysteine (NAC) improved the differentiation as well. We further found that ATP and NAC could partially replace the growth factor Activin A for definitive endoderm differentiation. Our study illustrates the essential role of mitochondria during human endoderm differentiation through providing ATP and regulating ROS levels, which may provide new insight for metabolic regulation of cell fate determination.

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