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

转铁蛋白 脂多糖 淋巴细胞 白蛋白 血清白蛋白 克隆(Java方法) 分泌物 生物 免疫学 化学 分子生物学 内科学 内分泌学 生物化学 医学 DNA
作者
Norman N. Iscove,Fritz Melchers
出处
期刊:Journal of Experimental Medicine [Rockefeller University Press]
卷期号:147 (3): 923-933 被引量:535
标识
DOI:10.1084/jem.147.3.923
摘要

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.

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