Hypoxic culture maintains self-renewal and enhances embryoid body formation of human embryonic stem cells

Hsin Fu Chen, Hung Chih Kuo, Shau Ping Lin, Chung Liang Chien, Ming Shan Chiang, Hong Nerng Ho

Research output: Contribution to journalArticle

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Abstract

Hypoxic environment is theoretically more physiological for the growth of human embryonic stem (hES) cells. It has been reported that hypoxic culture maintained better undifferentiation of hES cells, but the effects on differentiation are less well established. The hES cells were thus cultured and compared in hypoxia (2% oxygen [O2]) and normoxia (21% O 2). The data showed that the undifferentiated state of hES cells was maintained more favorably in hypoxia during prolonged culture. Most tested genes belonging to FGF, TGF-β/GMP, and Wnt signaling pathways were enriched in undifferentiated hES cells and downregulated upon differentiation, accompanied with differential expression of FGFR1, FGFR2, and FRAT2 between hypoxia and normoxia. Higher P-Smad2/3 level was identified in hypoxia, favoring the maintenance of hES cells in undifferentiation. Bisulfite sequencing showed similar imprinting status between different O2 tensions at H19 differentially methylated region (DMR) and KvDMR loci. Embryoid body formation was enhanced in hypoxia accompanied with suppressed Sox17, Desmin, Gata4, Brachyury, and Cdx2 expression. We concluded that hypoxia improved self-renewal of hES cells through modulation of major signaling pathways and was also more efficient for differentiation to embryoid bodies, though they might present with suppressed expression of some lineage-specific genes across all the three embryonic germ layers and trophectoderm.

Original languageEnglish
Pages (from-to)2901-2913
Number of pages13
JournalTissue Engineering - Part A
Volume16
Issue number9
DOIs
Publication statusPublished - Sep 1 2010

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T-Box Domain Proteins
SOXF Transcription Factors
Neoplasm Proteins
GATA4 Transcription Factor
Receptor, Fibroblast Growth Factor, Type 2
Receptor, Fibroblast Growth Factor, Type 1
Fetal Proteins
Embryoid Bodies
Homeodomain Proteins
Cell Hypoxia
Desmin
Fibroblast Growth Factors
Polymerase chain reaction
Embryonic Stem Cells
Stem cells
Gene expression
Transforming Growth Factor beta
Fluorescent Antibody Technique
Carrier Proteins
Stem Cells

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biomaterials
  • Biomedical Engineering

Cite this

Hypoxic culture maintains self-renewal and enhances embryoid body formation of human embryonic stem cells. / Chen, Hsin Fu; Kuo, Hung Chih; Lin, Shau Ping; Chien, Chung Liang; Chiang, Ming Shan; Ho, Hong Nerng.

In: Tissue Engineering - Part A, Vol. 16, No. 9, 01.09.2010, p. 2901-2913.

Research output: Contribution to journalArticle

Chen, Hsin Fu ; Kuo, Hung Chih ; Lin, Shau Ping ; Chien, Chung Liang ; Chiang, Ming Shan ; Ho, Hong Nerng. / Hypoxic culture maintains self-renewal and enhances embryoid body formation of human embryonic stem cells. In: Tissue Engineering - Part A. 2010 ; Vol. 16, No. 9. pp. 2901-2913.
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