Human lymphocyte transformation induced by mitogens and antigens in a serum-free tissue culture system |
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Authors: | Barbara White Needleman John M Weiler |
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Institution: | Department of Internal Medicine, University of Iowa, and the Iowa City Veterans Administration Hospital, Iowa City, IA 52242, U.S.A. |
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Abstract: | The use of serum to supplement lymphocyte tissue culture media introduces uncontrolled variables; different serum sources, lots and concentrations can produce variability in experimental results, serum can stimulate or inhibit lymphocytes, and components of serum can react with substances whose effects on lymphocytes are being studied. To avoid these problems, we studied the ability of human peripheral mononuclear cells to survive and to respond to stimulation in an entirely synthetic medium, RPMI-1640 supplemented with L-glutamine, gentamicin, HEPES buffer and magnesium. Optical cell concentration in this serum-free RPMI-1640 was 2.5 × 106 cells/ml, whereas optimal cell concentration in serum containing RPMI-1640 was 1 × 106 cells/ml. In this serum-free RPMI-1640, 50% of the cellular input was recovered as viable cells after 7 days of culture, which was similar to results in serum containing RPMI-1640. Mononuclear cell transformation was induced by phytohemagglutinin, concanavalin A, pokeweed mitogen, streptolysin O and candida. Optimal doses of stimulants and the kinetics of the responses were similar in serum-free and serum containing RPMI-1640. This system can be used to avoid the problems inherent in systems which supplement tissue culture media with serum. |
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Keywords: | Fellow of the Kidney Foundation of IowA Address requests for reprints: Dr Needleman 0080 Carver Pavilion University of Iowa Iowa City IA 52242 U S A |
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