Different murine-derived feeder cells alter the definitive endoderm differentiation of human induced pluripotent stem cells

Autor: Soichiro Ogaki, Yutaka Suzuki, Takashi Kuzuhara, Hiroki Minato, Masahide Seki, Masaki Shoji, Shoen Kume
Rok vydání: 2018
Předmět:
0301 basic medicine
Molecular biology
Cellular differentiation
lcsh:Medicine
Gene Expression
Stem cell marker
Biochemistry
Histones
Transcriptome
Mice
Sequencing techniques
lcsh:Science
Staining
Multidisciplinary
Transcriptional Control
Endoderm
Cell Staining
Cell Differentiation
RNA sequencing
Cell biology
KLF4
embryonic structures
Signal Transduction
Research Article
Homeobox protein NANOG
Induced Pluripotent Stem Cells
Biology
Cell Line
Kruppel-Like Factor 4
03 medical and health sciences
Extraction techniques
SOX2
DNA-binding proteins
Genetics
Animals
Humans
Gene Regulation
lcsh:R
Feeder Cells
Biology and Life Sciences
Proteins
Fibroblasts
Embryonic stem cell
Coculture Techniques
RNA extraction
Research and analysis methods
Molecular biology techniques
030104 developmental biology
Specimen Preparation and Treatment
Cell culture
lcsh:Q
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 13, Iss 7, p e0201239 (2018)
ISSN: 1932-6203
Popis: The crosstalk between cells is important for differentiation of cells. Murine-derived feeder cells, SNL76/7 feeder cells (SNLs) or mouse primary embryonic fibroblast feeder cells (MEFs) are widely used for culturing undifferentiated human induced pluripotent stem cells (hiPSCs). It is still unclear whether different culture conditions affect the induction efficiency of definitive endoderm (DE) differentiation from hiPSCs. Here we show that the efficiency of DE differentiation from hiPSCs cultured on MEFs was higher than that of hiPSCs cultured on SNLs. The qPCR, immunofluorescent and flow cytometry analyses revealed that the expression levels of mRNA and/or proteins of the DE marker genes, SOX17, FOXA2 and CXCR4, in DE cells differentiated from hiPSCs cultured on MEFs were significantly higher than those cultured on SNLs. Comprehensive RNA sequencing and molecular network analyses showed the alteration of the gene expression and the signal transduction of hiPSCs cultured on SNLs and MEFs. Interestingly, the expression of non-coding hXIST exon 4 was up-regulated in hiPSCs cultured on MEFs, in comparison to that in hiPSCs cultured on SNLs. By qPCR analysis, the mRNA expression of undifferentiated stem cell markers KLF4, KLF5, OCT3/4, SOX2, NANOG, UTF1, and GRB7 were lower, while that of hXIST exon 4, LEFTY1, and LEFTY2 was higher in hiPSCs cultured on MEFs than in those cultured on SNLs. Taken together, our finding indicated that differences in murine-feeder cells used for maintenance of the undifferentiated state alter the expression of pluripotency-related genes in hiPSCs by the signaling pathways and affect DE differentiation from hiPSCs, suggesting that the feeder cells can potentiate hiPSCs for DE differentiation.
Databáze: OpenAIRE
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