Serum increases the CD13 receptor expression, reduces the transduction of fluid-mechanical forces, and alters the metabolism of HL60 cells cultured in agitated bioreactors.
The effects of serum medium concentration on the CD13 receptor surface content and mRNA levels of HL60 (human promyelocytic leukemia) cells were examined using flow cytometry and Northern blotting. Increasing the serum concentration from 2.5% to 10% and from 5% to 10% increased the CD13 receptor surface content of HL60 cells by 100% and 25%, respectively, in spinner flasks agitated at 60 rpm. In bioreactors at 80 rpm, increasing the serum concentration from 2.5% to 10% and from 5% to 10% increased the CD13 receptor surface content by 60% and 35%, respectively. This increase in CD13 receptor surface content was correlated with a 30% and a 20% increase in CD13 mRNA levels. Increasing serum concentrations also increased the average HL60 cell size under non-damaging conditions (60 rpm in spinner flasks, 80 rpm in bioreactors). Under conditions of agitation at 300 rpm in 2 L bioreactors, increasing serum concentrations (2.5% vs. 10%, 5% vs. 10%) allowed for higher HL60 apparent growth rates, but decreased the CD13 receptor surface content and mRNA levels. In view of our earlier findings on the effects of agitation on the CD13 antigen, these data suggest that serum reduces the transduction of mechanical forces that affect CD13 expression. At 300 rpm, HL60 cells cultured in 10% serum exhibited glucose consumption and lactate production rates that were approximately 50% and 60% lower than the values of cells cultured in 5% and 2.5% serum, respectively.
McDowell
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Northwestern University, Department of Chemical Engineering, 2145 Sheridan Road, Evanston, Illinois 60208-3120, USA.
Papoutsakis
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D019149:Bioreactors; D001769:Blood; D018826:CD13 Antigens / Q000096:biosynthesis / Q000235:genetics*; D018929:Cell Culture Techniques / Q000379:methods; D003470:Culture Media; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism; D018922:HL-60 Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D007773:Lactates / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D013314:Stress, Mechanical; D014158:Transcription, Genetic
D003470:Culture Media; D007773:Lactates; D012333:RNA, Messenger; D018826:CD13 Antigens; D005947:Glucose