Metabolic phenotyping to identify cellular transitions during cardiomyocytes differentiation from human embryonic stem cells
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1.1)School of Life Science, Beijing Institute of Technology, Beijing 100081, China;2.2)Center of Biomedical Analysis, Tsinghua University, Beijing 100084, China

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National 863 Program in China(2013AA032204)

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    Abstract:

    Objective :We performed directional differentiation of human embryonic stem cells into cardiomyocytes. In order to explore the mechanisms of cell metabolic phenotype conversion during cardiac lineage differentiation ,we conducted real-time quantitative detection of glycolytic and mitochondrial oxidative phosphorylation capabilities of embryonic stem cells, cardiac progenitor cells, and cardiomyocytes during differentiation. Methods: GSK3 inhibitor CHIR99021 and Wnt signaling pathway inhibitor IWP2 were used to differentiate human embryonic stem cells into cardiac progenitor cells and cardiomyocytes. Immunocytochemistry was used to detect the expression of human embryonic stem cell markers. Flow cytometry was used to detect the markers of human cardiomyocytes and cardiac progenitor cells. Extracellular Flux Analysis was used to test the energy metabolic phenotype of human embryonic stem cells, cardiac progenitor cells, and cardiomyocytes. Results: The stemness of human embryonic stem cells remains stable and all express Nanog, OCT4 and SOX2 cell markers. During the differentiation, more than 99% cells expressed cardiac progenitor cell marker Isl1 on the 7th day, and more than 83% of cells expressed the cardiomyocytes marker cTnT on the 15th day. Human embryonic stem cells have the strongest glycolytic metabolism capacity, while cardiomyocytes have the strongest mitochondrial oxidative phosphorylation capability. Cardiac progenitor cells are in the transition stage of the two ways of metabolism. Conclusion: During the differentiation of human embryonic stem cells into cardiomyocytes, cells gradually loss the glycolytic capacity, while the mitochondrial oxidative phosphorylation capacity gradually increases, followed by the cell metabolic phenotype conversion.

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JIAO Peng-Cheng, JI Jiao-Jiao, ZHAO Dong-xu. Metabolic phenotyping to identify cellular transitions during cardiomyocytes differentiation from human embryonic stem cells[J]. Progress in Biochemistry and Biophysics,2020,47(11):1191-1198

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History
  • Received:April 16,2020
  • Revised:June 06,2020
  • Accepted:June 12,2020
  • Online: March 04,2021
  • Published: November 20,2020