Increased agitation intensity increases CD13 receptor surface content and mRNA levels, and alters the metabolism of HL60 cells cultured in stirred tank bioreactors.
Flow cytometry and Northern blotting were used to examine the effects of hydrodynamic forces in stirred tank bioreactors on CD13 receptor surface content and mRNA levels of HL60 (human promyelocytic leukemia) cells. A step increase in agitation rate from 80 to 300 or 400 rpm reduced the apparent HL60 growth rate in a dose-dependent manner. This step increase in agitation rate (to 300 or 400 rpm) also increased the CD13 receptor surface content on averge by 30% and 100%, respectively. This increase in CD13 receptor surface content was correlated with a 10% and a 60% increase in CD13 mRNA levels. We also observed a significant and very reproducible drop in CD13 expression over the course of a batch bioreactor run (80 rpm). Although we have no explanation for this, we show that the decrease in CD13 receptor surface content can be (at least partially, if not fully) explained by the corresponding decrease in CD13 mRNA. HL60 cell cultures agitated at 300 and 400 rpm exhibited glucose consumption and lactate production rates that were approximately 40% and 90% greater than values of the cultures agitated at 80 rpm. The physiological and practical implications of these results are discussed.
McDowell
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Northwestern University, Department of Chemical Engineering, 2145 Sheridan Road, Evanston, Illinois 60208-3120, USA.
Papoutsakis
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D000954:Antigens, Surface / Q000096:biosynthesis / Q000235:genetics; D019149:Bioreactors; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D018826:CD13 Antigens / Q000096:biosynthesis / Q000235:genetics*; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism; D018922:HL-60 Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics
D000954:Antigens, Surface; D012333:RNA, Messenger; D018826:CD13 Antigens; D005947:Glucose