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
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DOI：
10.1084/jem.147.3.923
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发表时间：
1978-01-01
影响因子：
15.3
通讯作者：
MELCHERS, F
中科院分区：
文献类型：
--
作者：
ISCOVE, NN;MELCHERS, F
白蛋白、转铁蛋白和脂质可完全替代血清，以支持培养物中LPS[脂多糖]刺激的小鼠B [骨髓来源]淋巴细胞。在这些化合物的存在下，在无血清条件下，甚至在相对低的细胞浓度下，由LPS诱导的生长和成熟至IG[免疫球蛋白]M和IgG分泌的效率与在常规含血清培养基中相同或更高。与常规含血清培养基中LPS反应性的快速消失相反，在无血清条件下反应性保持在初始水平2天，然后缓慢下降。淋巴细胞的总体存活时间也显着延长。在存在胸腺填充细胞的情况下，无血清条件允许每种LPS应答细胞生长至低至单个反应性B细胞/培养物的稀释度的Ig分泌细胞克隆。研究结果有几个重要的意义。其中包括首次将转铁蛋白确定为培养物中B淋巴细胞反应的必要条件，以及现在已具备测定和分离调节淋巴细胞功能的细胞产物的条件，且不受不确定血清成分的干扰。
Albumin, transferrin and lipids can replace serum entirely for support of LPS[lipopolysaccharide]-stimulated murine B [bone marrow-derived] lymphocytes in culture. In the presence of these compounds, growth and maturation to Ig[immunoglobulin]M and IgG secretion, induced by 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/culture. The results have several important implications. These include the establishment for the 1st time of transferrin as a requirement for B lymphocyte responses in culture, and the availability now of conditions for the assay and isolation of cell products regulating lymphocyte function, free of interference from undefined serum components.