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在脂多糖反应性B淋巴细胞培养物中用白蛋白、转铁蛋白和大豆脂质完全替代血清。

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

作者信息

Iscove N N, Melchers F

出版信息

J Exp Med. 1978 Mar 1;147(3):923-33. doi: 10.1084/jem.147.3.923.

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.

摘要

白蛋白、转铁蛋白和脂质可以完全替代血清,以支持培养中脂多糖刺激的小鼠B淋巴细胞。在这些化合物存在的情况下,脂多糖(LPS)诱导的生长以及向IgM和IgG分泌的成熟过程,在无血清条件下与在传统含血清培养基中以相同或更高的效率发生,即使在相对较低的细胞浓度下也是如此。与传统含血清培养基中LPS反应性的快速消失相反,在无血清条件下,反应性在缓慢下降之前的2天内保持在初始水平。总体淋巴细胞存活率也显著延长。在胸腺“填充”细胞存在的情况下,无血清条件允许每个LPS反应性细胞生长为一个分泌Ig的细胞克隆,稀释度低至每培养物一个反应性B细胞。这些结果有几个重要意义。其中包括首次确定转铁蛋白是培养中B淋巴细胞反应的必需物质,以及现在有了用于检测分离调节淋巴细胞功能的细胞产物的条件,且不受未定义血清成分的干扰。

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