RRC ID 61177
Author Tomizawa M, Shinozaki F, Motoyoshi Y, Sugiyama T, Yamamoto S, Ishige N.
Title 2‑Deoxy‑D‑glucose initiates hepatocyte differentiation in human induced pluripotent stem cells.
Journal Mol Med Rep
Abstract To initiate hepatocyte differentiation in human induced pluripotent stem (iPS) cells, cells are cultured in a medium lacking glucose but supplemented with galactose (hepatocyte selection medium, HSM) or in medium supplemented with oncostatin M and small molecules (hepatocyte differentiation inducer, HDI). In the present study, 2‑Deoxy‑D‑glucose (2DG), an analogue of glucose, was utilized instead of glucose deprivation and the effect of 2DG supplementation on iPS differentiation was examined. First, 201B7 cells, an iPS cell line, were cultured in HSM or HDI media for 2 days and then subjected to reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) in order to analyze expression levels of established hepatocyte markers, including cytosolic aspartate aminotransferase (AST), mitochondrial AST, alanine aminotransferase (ALT), and glycerol kinase. mRNA expression levels of mitochondrial AST, ALT, and glycogen synthase significantly increased following culture in HSM and HDI compared with ReproFF media. Cytosolic AST mRNA expression levels significantly increased following culture in HDI compared with ReproFF media, but not in HSM. To test the effect of 2DG on iPS differentiation, 201B7 cells were cultured in ReproFF, a feeder‑free medium that retains pluripotency, supplemented with 2DG. Following 7 days of culture, the cells were subjected to RT‑qPCR to analyze expression levels of α‑fetoprotein (AFP), a marker of immature hepatocytes. AFP mRNA expression levels significantly increased with the addition of 0.1 µM 2DG in the media, and galactose addition acted synergistically with 2DG to further upregulate AFP expression. In conclusion, the present study demonstrated that hepatocyte differentiation was initiated in iPS cells cultured in HSM and HDI media and that 2DG could be used as a supplement instead of glucose deprivation to initiate hepatocyte differentiation in iPS cells.
Volume 15(5)
Pages 3083-3087
Published 2017-5-1
DOI 10.3892/mmr.2017.6405
PMID 28358426
MeSH Alanine Transaminase / metabolism Aspartate Aminotransferase, Cytoplasmic / metabolism Aspartate Aminotransferase, Mitochondrial / metabolism Cell Differentiation / drug effects* Cell Line Deoxyglucose / pharmacology* Galactose / metabolism Glucose / metabolism Glycerol Kinase / metabolism Glycogen Synthase / metabolism Hepatocytes / cytology* Hepatocytes / drug effects* Hepatocytes / metabolism Humans Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / drug effects* Induced Pluripotent Stem Cells / metabolism Oncostatin M / pharmacology RNA, Messenger / metabolism Real-Time Polymerase Chain Reaction / methods alpha-Fetoproteins / metabolism
IF 2.1
Human and Animal Cells 201B7(HPS0063)