A reduced oxygen tension (5%) is not beneficial for maintaining human embryonic stem cells in the undifferentiated state with short splitting intervals

Hsin Fu Chen, Hung Chih Kuo, Wannhsin Chen, Fang Chun Wu, Yu Shih Yang, Hong Nerng Ho

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

BACKGROUND: Human embryos grow naturally in vivo in lower oxygen (O 2) tension environments than atmospheric O2 tension. Therefore, human embryonic stem cells (hESC), a derivative of embryos, will likely grow more favorably in a reduced O2 tension. This study aimed to compare the behavior of hESC under reduced O2 tension (5%) versus normoxia (21%). METHODS: hESC lines were cultured in different O2 tensions and then examined for morphology, apoptosis and gene expression profiles. RESULTS: hESC grown in 5% O2 tension were not morphologically different from hESC grown in normoxia on day 7 of the first and fourth passages. However, after prolonged culture without splitting (10-14 days), hESC colonies were thinner and looked better morphologically in 5% O 2, but the cells proliferated more slowly and their sizes were larger. At most time points, the gene expression profiles in both O2 tensions showed no major difference in representative stemness genes (Oct-3/4, Nanog and Cripto), differentiation genes (Desmin, Nestin, α-fetoprotein and GDF-9) and hypoxia-related genes (HIF-1α and VEGF). A lower level of cyclin-D1 mRNA (suggestive of less Wnt pathway signaling on day 7 of the fourth passage) and a higher level of Desmin (suggestive of more differentiation to mesoderm, at day 7 of the first passage) were detected in 5% O2. CONCLUSIONS: This study suggests that for routine culture of hESC with a short splitting interval (7 days), a low O2 tension (5% O2) probably does not provide significant advantages over the standard 21% O 2 tension for the maintenance of an undifferentiated state by the criteria used in this study.

Original languageEnglish
Pages (from-to)71-80
Number of pages10
JournalHuman Reproduction
Volume24
Issue number1
DOIs
Publication statusPublished - Jan 1 2009
Externally publishedYes

Fingerprint

Octamer Transcription Factor-3
Wnt Proteins
Cell Hypoxia
Gene Expression Profiling
Proteoglycans
Surface Antigens
Embryonic Stem Cells
Alkaline Phosphatase
Cell Differentiation
Signal Transduction
Cell Culture Techniques
Cell Proliferation
Apoptosis
Oxygen
Cell Line
Desmin
Transcriptome
Growth Differentiation Factor 9
Embryonic Structures
Genes

Keywords

  • Cell culture
  • Embryonic stem cells
  • Hypoxia
  • Oxygen
  • Undifferentiated

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynaecology

Cite this

A reduced oxygen tension (5%) is not beneficial for maintaining human embryonic stem cells in the undifferentiated state with short splitting intervals. / Chen, Hsin Fu; Kuo, Hung Chih; Chen, Wannhsin; Wu, Fang Chun; Yang, Yu Shih; Ho, Hong Nerng.

In: Human Reproduction, Vol. 24, No. 1, 01.01.2009, p. 71-80.

Research output: Contribution to journalArticle

Chen, Hsin Fu ; Kuo, Hung Chih ; Chen, Wannhsin ; Wu, Fang Chun ; Yang, Yu Shih ; Ho, Hong Nerng. / A reduced oxygen tension (5%) is not beneficial for maintaining human embryonic stem cells in the undifferentiated state with short splitting intervals. In: Human Reproduction. 2009 ; Vol. 24, No. 1. pp. 71-80.
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abstract = "BACKGROUND: Human embryos grow naturally in vivo in lower oxygen (O 2) tension environments than atmospheric O2 tension. Therefore, human embryonic stem cells (hESC), a derivative of embryos, will likely grow more favorably in a reduced O2 tension. This study aimed to compare the behavior of hESC under reduced O2 tension (5{\%}) versus normoxia (21{\%}). METHODS: hESC lines were cultured in different O2 tensions and then examined for morphology, apoptosis and gene expression profiles. RESULTS: hESC grown in 5{\%} O2 tension were not morphologically different from hESC grown in normoxia on day 7 of the first and fourth passages. However, after prolonged culture without splitting (10-14 days), hESC colonies were thinner and looked better morphologically in 5{\%} O 2, but the cells proliferated more slowly and their sizes were larger. At most time points, the gene expression profiles in both O2 tensions showed no major difference in representative stemness genes (Oct-3/4, Nanog and Cripto), differentiation genes (Desmin, Nestin, α-fetoprotein and GDF-9) and hypoxia-related genes (HIF-1α and VEGF). A lower level of cyclin-D1 mRNA (suggestive of less Wnt pathway signaling on day 7 of the fourth passage) and a higher level of Desmin (suggestive of more differentiation to mesoderm, at day 7 of the first passage) were detected in 5{\%} O2. CONCLUSIONS: This study suggests that for routine culture of hESC with a short splitting interval (7 days), a low O2 tension (5{\%} O2) probably does not provide significant advantages over the standard 21{\%} O 2 tension for the maintenance of an undifferentiated state by the criteria used in this study.",
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T1 - A reduced oxygen tension (5%) is not beneficial for maintaining human embryonic stem cells in the undifferentiated state with short splitting intervals

AU - Chen, Hsin Fu

AU - Kuo, Hung Chih

AU - Chen, Wannhsin

AU - Wu, Fang Chun

AU - Yang, Yu Shih

AU - Ho, Hong Nerng

PY - 2009/1/1

Y1 - 2009/1/1

N2 - BACKGROUND: Human embryos grow naturally in vivo in lower oxygen (O 2) tension environments than atmospheric O2 tension. Therefore, human embryonic stem cells (hESC), a derivative of embryos, will likely grow more favorably in a reduced O2 tension. This study aimed to compare the behavior of hESC under reduced O2 tension (5%) versus normoxia (21%). METHODS: hESC lines were cultured in different O2 tensions and then examined for morphology, apoptosis and gene expression profiles. RESULTS: hESC grown in 5% O2 tension were not morphologically different from hESC grown in normoxia on day 7 of the first and fourth passages. However, after prolonged culture without splitting (10-14 days), hESC colonies were thinner and looked better morphologically in 5% O 2, but the cells proliferated more slowly and their sizes were larger. At most time points, the gene expression profiles in both O2 tensions showed no major difference in representative stemness genes (Oct-3/4, Nanog and Cripto), differentiation genes (Desmin, Nestin, α-fetoprotein and GDF-9) and hypoxia-related genes (HIF-1α and VEGF). A lower level of cyclin-D1 mRNA (suggestive of less Wnt pathway signaling on day 7 of the fourth passage) and a higher level of Desmin (suggestive of more differentiation to mesoderm, at day 7 of the first passage) were detected in 5% O2. CONCLUSIONS: This study suggests that for routine culture of hESC with a short splitting interval (7 days), a low O2 tension (5% O2) probably does not provide significant advantages over the standard 21% O 2 tension for the maintenance of an undifferentiated state by the criteria used in this study.

AB - BACKGROUND: Human embryos grow naturally in vivo in lower oxygen (O 2) tension environments than atmospheric O2 tension. Therefore, human embryonic stem cells (hESC), a derivative of embryos, will likely grow more favorably in a reduced O2 tension. This study aimed to compare the behavior of hESC under reduced O2 tension (5%) versus normoxia (21%). METHODS: hESC lines were cultured in different O2 tensions and then examined for morphology, apoptosis and gene expression profiles. RESULTS: hESC grown in 5% O2 tension were not morphologically different from hESC grown in normoxia on day 7 of the first and fourth passages. However, after prolonged culture without splitting (10-14 days), hESC colonies were thinner and looked better morphologically in 5% O 2, but the cells proliferated more slowly and their sizes were larger. At most time points, the gene expression profiles in both O2 tensions showed no major difference in representative stemness genes (Oct-3/4, Nanog and Cripto), differentiation genes (Desmin, Nestin, α-fetoprotein and GDF-9) and hypoxia-related genes (HIF-1α and VEGF). A lower level of cyclin-D1 mRNA (suggestive of less Wnt pathway signaling on day 7 of the fourth passage) and a higher level of Desmin (suggestive of more differentiation to mesoderm, at day 7 of the first passage) were detected in 5% O2. CONCLUSIONS: This study suggests that for routine culture of hESC with a short splitting interval (7 days), a low O2 tension (5% O2) probably does not provide significant advantages over the standard 21% O 2 tension for the maintenance of an undifferentiated state by the criteria used in this study.

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KW - Embryonic stem cells

KW - Hypoxia

KW - Oxygen

KW - Undifferentiated

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