Effect of oxygen on the growth of human epidermal keratinocytes.
We studied the growth of secondary cultures of neonatal human keratinocytes at oxygen concentrations between 1 and 89%. Keratinocytes were grown in MCDB medium with 5% fetal bovine serum, 10 ng/ml epidermal growth factor, 5 micrograms/ml insulin, 0.5 microgram/ml hydrocortisone, and 0.1 mM ethanolamine and phosphorylethanolamine. Medium in the flasks was equilibrated with gas mixtures containing 5% CO2, various percentages of oxygen from 0-95% and nitrogen to balance. Cells were seeded at 10(4) cells/cm2 in sealed flasks (25 cm2). These were incubated at 37 degrees C in incubators maintained at the experimental oxygen tensions. Cells grew best at PO2 (partial pressure of oxygen) 133 mm Hg (18% O2), with a mean population doubling time of 2.8 days. Growth was retarded by 60% at PO2 38 mm Hg (5% O2) and by 98% at PO2 7 mm Hg (1% O2). However, the oxygen tension that resulted in the best plating efficiency was at PO2 12 mm Hg (2% O2). When oxygen tensions were shifted to 78-133 mm Hg, cells seeded under low oxygen tensions began to proliferate. These data suggest that a better harvest of keratinocytes is obtained when cells are seeded under low oxygen tension and then shifted to ambient oxygen tensions. At high oxygen tensions, above 20%, growth was inhibited by 75% at PO2 241 mm Hg (34% O2) and 98% at PO2 374 mm Hg (52% O2). At PO2 637 mm Hg (89% O2) no cell growth occurred. These findings showed that high oxygen tensions, above 20%, have no beneficial effect on the growth of keratinocytes.
D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects; D006801:Humans; D007231:Infant, Newborn; D007633:Keratins; D010100:Oxygen / Q000494:pharmacology*; D010313:Partial Pressure
D003470:Culture Media; D007633:Keratins; D010100:Oxygen