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Transcription Factors and Medium Suitable for Initiating the Differentiation of Human‐Induced Pluripotent Stem Cells to the Hepatocyte Lineage
Author(s) -
Tomizawa Minoru,
Shinozaki Fuminobu,
Motoyoshi Yasufumi,
Sugiyama Takao,
Yamamoto Shigenori,
Ishige Naoki
Publication year - 2016
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25494
Subject(s) - cebpa , biology , foxa2 , foxa1 , microbiology and biotechnology , induced pluripotent stem cell , transfection , transcription factor , ccaat enhancer binding proteins , homeobox protein nanog , enhancer , stem cell , cell culture , gene , embryonic stem cell , genetics , dna binding protein
Transcription factors and culture media were investigated to determine the condition to initiate the differentiation of human‐induced pluripotent stem (iPS) cells most efficiently. The expression of genes in human adult liver was compared with that in 201B7 cells (iPS cells) using cDNA microarray analysis. Episomal plasmids expressing transcription factors were constructed. 201B7 cells were transfected with the episomal plasmids and cultured in ReproFF (feeder‐free media maintaining pluripotency), Leibovitz‐15 (L15), William's E (WE), or Dulbecco's modified Eagle medium/Nutrient F‐12 Ham (DF12) for 7 days. RNA was isolated and subjected to real‐time quantitative PCR to analyze the expression of alpha‐feto protein (AFP) and albumin. cDNA microarray analysis revealed 16 transcription factors that were upregulated in human adult liver relative to that in 201B7 cells. Episomal plasmids expressing these 16 genes were transfected into 201B7 cells. CCAAT/enhancer‐binding protein alpha (CEBPA), CCAAT/enhancer‐binding protein beta (CEBPB), forkhead box A1 (FOXA1), and forkhead box A3 (FOXA3) up‐regulated AFP and down‐regulated Nanog. These four genes were further analyzed. The expression of AFP and albumin was the highest in 201B7 cells transfected with the combination of CEBPA, CEBPB, FOXA1, and FOXA3 and cultured in WE. The combination of CEBPA, CEBPB, FOXA1, and FOXA3 was suitable for 201B7 cells to initiate differentiation to the hepatocyte lineage and WE was the most suitable medium for culture after transfection. J. Cell. Biochem. 117: 2001–2009, 2016. © 2016 Wiley Periodicals, Inc.