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  • Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes.

Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes.

The Journal of experimental medicine (1978-03-01)
N N Iscove, F Melchers
PMID305462
ABSTRACT

Albumin, transferrin, and lipids can replace serum entirely for support of LPS-stimulated murine B lymphocytes in culture. In the presence of these compounds, growth and maturation to IgM and IgG secretion, induced by lipopolysaccharide (LPS), occurs at the same or higher efficiency in serum-free conditions as in conventional serum-containing medium, even at relatively low cell concentrations. In contrast to the rapid disappearance of LPS reactivity in conventional serum-containing medium, responsiveness remains at initial levels in serum-free conditions for 2 days before slowly declining. Overall lymphocyte survival is also markedly prolonged. In the presence of thymus "filler" cells, the serum-free conditions permit growth of every LPS-responsive cell to a clone of Ig-secreting cells at dilutions as low as a single reactive B cell per culture. The results have several important implications. These include the establishment for the first time of transferrin as a requirement for B lymphocyte responses in culture, and the availability now of conditions for the assay isolation of cell products regulating lymphocyte function, free of interference from undefined serum components.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Iscove′s Modified Dulbecco′s Medium, liquid, sterile-filtered, With sodium bicarbonate, without L-glutamine, suitable for cell culture, suitable for hybridoma