The stromal cell line S17 supports the growth of lipopolysaccharide-stimulated CBA/N spleen cell coloniesin vitro
- 1 April 1992
- journal article
- research article
- Published by Wiley in European Journal of Immunology
- Vol. 22 (4) , 1001-1006
- https://doi.org/10.1002/eji.1830220419
Abstract
The expression of an X-linked defect in the CBA/N strain of mice has been found to result in a number of immune abnormalities. These include low responsiveness to antigens and a greatly reduced ability to respond to many of the common B cell mitogens. An in vitro manifestation of this condition is the virtual inability of CBA/N B cells to form colonies in lipopolysaccharide (LPS)-containing semisolid agar cultures. In this report we show evidence that the colony-forming ability of CBA/N spleen cells can be effectively restored by the bone marrow stromalderived cell line S17. Spleen cells from 4–5-week-old homozygous CBA/N female mice were grown in double-layer agar cultures containing S17 feeder layers. Control cultures contained the fibroblast-like cell line 95.17 or were treated with medium alone. It was found that at an input cell concentration of 104 cells per plate, CBA/N colony formation was increased from a frequency of approximately 1 in 5000 to 1 in 50 total splenic cells. Studies with purified surface immunoglobulin-positive cells indicate the direct involvement of S17 in this process. The CBA/N colonies formed were dependent on the presence of a mitogen (LPS) and secreted detectable amounts of IgM. Major implications of these findings and the application of this assay system to study the CBA/N defect have been discussed.Keywords
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