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Effects of oxygen on mouse embryonic stem cell growth, phenotype retention, and cellular energetics
Author(s) -
Powers Daryl E.,
Millman Jeffrey R.,
Huang Ryan B.,
Colton Clark K.
Publication year - 2008
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21986
Subject(s) - leukemia inhibitory factor , oxygen , embryonic stem cell , cellular differentiation , stem cell , chemistry , metabolism , biology , cell growth , microbiology and biotechnology , cell , anaerobic exercise , hypoxia (environmental) , biochemistry , physiology , organic chemistry , gene
Abstract Most embryonic stem (ES) cell research is performed with a gas phase oxygen partial pressure (pO 2 ) of 142 mmHg, whereas embryonic cells in early development are exposed to pO 2 values of 0–30 mmHg. To understand effects of these differences, we studied murine ES (mES) growth, maintenance of stem cell phenotype, and cell energetics over a pO 2 range of 0–285 mmHg, in the presence or absence of differentiation‐suppressing leukemia inhibitory factor (LIF). With LIF, growth rate was sensitive to pO 2 but constant with time, and expression of self‐renewal transcription factors decreased at extremes of pO 2 . Subtle morphological changes suggested some early differentiation, but cells retained the ability to differentiate into derivatives of all three germ layers at low pO 2 . Without LIF, growth rate decreased with time, and self‐renewal transcription factor mRNA decreased further. Gross morphological changes occurred, and overt differentiation occurred at all pO 2 . These findings suggested that hypoxia in the presence of LIF promoted limited early differentiation. ES cells survived oxygen starvation with negligible cell death by increasing anaerobic metabolism within 48 h of anoxic exposure. Decreasing pO 2 to 36 mmHg or lower decreased oxygen consumption rate and increased lactate production rate. The fraction of ATP generated aerobically was 60% at or above 142 mmHg and decreased to 0% under anoxia, but the total ATP production rate remained nearly constant at all pO 2 . In conclusion, undifferentiated ES cells adapt their energy metabolism to proliferate at all pO 2 between 0 and 285 mmHg. Oxygen has minimal effects on undifferentiated cell growth and phenotype, but may exert more substantial effects under differentiating conditions. Biotechnol. Bioeng. 2008;101: 241–254. © 2008 Wiley Periodicals, Inc.