Effects of methocel A15LV, polyethylene glycol, and polyvinyl alcohol on CD13 and CD33 receptor surface content and metabolism of HL60 cells cultured in stirred tank bioreactors.
Flow cytometry was used to examine the effect of hydrodynamic forces in a stirred tank bioreactor on the CD13 and CD33 receptor surface content of HL60 (human promyelocytic leukemia) cells. A step increase in agitation rate from 80 to 400 rpm reduced the HL60 cell apparent growth rate and increased the CD13 receptor surface content per cell, on average, by 95%. In contrast, this step increase in agitation rate to 400 rpm decreased the CD33 receptor surface content per cell, on average, by 10%. The protective effects of 0.1% Methocel A15LV, polyethylene glycol (PEG), and polyvinyl alcohol (PVA) on CD13 and CD33 receptor surface content were examined under agitation at 300 rpm in parallel 2 L bioreactor runs. The average CD33 receptor surface content was unaffected by the presence of Methocel A15LV or PEG, while PVA had a slight protective effect. In contrast, in terms of CD13 receptor content, HL60 cells agitated at 300 rpm with Methocel A15LV, PEG, or PVA behaved like cells agitated at 80 rpm with no media additives (McDowell and Papoutsakis, 1998). That is, Methocel A15LV, PEG, and PVA prevented the transduction of mechanical forces which affect CD13 cell content. HL60 cells cultured with 0.1% A15LV, PEG or PVA under conditions of mild agitation (60 rpm) in spinner flasks exhibited glucose consumption and lactate production rates that were approximately 20% lower than values of cultures containing no additive. Under conditions of agitation at 300 rpm in the 2 L bioreactor, the presence of A15LV, PEG, and PVA reduced the HL60 glucose consumption and lactate production rates by approximately 50%. Thus, media additives can dramatically reduce lactate accumulation in agitated bioreactors due to cell growth, in addition to providing protection from cellular injury.
McDowell
C L
CL
Northwestern University, Department of Chemical Engineering, 2145 Sheridan Road, Evanston, Illinois 60208-3120, USA.
Carver
R T
RT
Papoutsakis
E T
ET
D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics; D015214:Antigens, Differentiation, Myelomonocytic / Q000096:biosynthesis* / Q000235:genetics; D019149:Bioreactors; D018826:CD13 Antigens / Q000096:biosynthesis* / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods; D005947:Glucose / Q000378:metabolism; D018922:HL-60 Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D007773:Lactates / Q000378:metabolism; D008747:Methylcellulose / Q000494:pharmacology*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011142:Polyvinyl Alcohol / Q000494:pharmacology*; D063268:Sialic Acid Binding Ig-like Lectin 3
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; C571163:CD33 protein, human; D007773:Lactates; D063268:Sialic Acid Binding Ig-like Lectin 3; D011092:Polyethylene Glycols; D011142:Polyvinyl Alcohol; D008747:Methylcellulose; D018826:CD13 Antigens; D005947:Glucose