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Energy Metabolism in Human Pluripotent Stem and Differentiated Cells Compared Using a Seahorse XF96 Extracellular Flux Analyzer.
Kim, Hyun Kyu; Song, Yena; Kye, Minji; Yu, Byeongho; Park, Sang Beom; Kim, Ji Hyeon; Moon, Sung-Hwan; Choi, Hyungkyu; Moon, Jong-Seok; Oh, Jae Sang; Lee, Man Ryul.
Afiliación
  • Kim HK; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Song Y; Dementia Research Group, Korea Brain Research Institute (KBRI), Daegu, Korea.
  • Kye M; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Yu B; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Park SB; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Kim JH; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Moon SH; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Choi H; Department of Animal Science and Technology College of Biotechnology, Chung-Ang University, Anseong, Korea.
  • Moon JS; Department of Animal Science and Technology College of Biotechnology, Chung-Ang University, Anseong, Korea.
  • Oh JS; Soonchunhyang Institute of Medi-bio Science (SIMS), Soon Chun Hyang University, Cheonan, Korea.
  • Lee MR; Department of Neurosurgery, Uijeonbu St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea.
Int J Stem Cells ; 17(2): 194-203, 2024 May 30.
Article en En | MEDLINE | ID: mdl-38664993
ABSTRACT
Evaluating cell metabolism is crucial during pluripotent stem cell (PSC) differentiation and somatic cell reprogramming as it affects cell fate. As cultured stem cells are heterogeneous, a comparative analysis of relative metabolism using existing metabolic analysis methods is difficult, resulting in inaccuracies. In this study, we measured human PSC basal metabolic levels using a Seahorse analyzer. We used fibroblasts, human induced PSCs, and human embryonic stem cells to monitor changes in basal metabolic levels according to cell number and determine the number of cells suitable for analysis. We evaluated normalization methods using glucose and selected the most suitable for the metabolic analysis of heterogeneous PSCs during the reprogramming stage. The response of fibroblasts to glucose increased with starvation time, with oxygen consumption rate and extracellular acidification rate responding most effectively to glucose 4 hours after starvation and declining after 5 hours of starvation. Fibroblasts and PSCs achieved appropriate responses to glucose without damaging their metabolism 2∼4 and 2∼3 hours after starvation, respectively. We developed a novel method for comparing basal metabolic rates of fibroblasts and PSCs, focusing on quantitative analysis of glycolysis and oxidative phosphorylation using glucose without enzyme inhibitors. This protocol enables efficient comparison of energy metabolism among cell types, including undifferentiated PSCs, differentiated cells, and cells undergoing cellular reprogramming, and addresses critical issues, such as differences in basal metabolic levels and sensitivity to normalization, providing valuable insights into cellular energetics.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Int J Stem Cells Año: 2024 Tipo del documento: Article Pais de publicación: Corea del Sur
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Int J Stem Cells Año: 2024 Tipo del documento: Article Pais de publicación: Corea del Sur