Jun 1, 1989

Detection of interleukin 1 with human dermal fibroblasts

Immunobiology
H LoppnowE Brandt

Abstract

A fibroblast proliferation assay was developed for the detection of interleukin 1 (IL 1). Proliferation was measured by thymidine incorporation and by staining of cellular proteins with crystal violet. Response of fibroblasts was optimal at cell numbers of 4,000 to 9,000 cells/culture and an incubation period of four days. Serum content of the culture medium, ranging from 1 to 10% fetal calf serum (FCS), enhanced the proliferative response in a concentration-dependent manner, while higher concentrations of FCS did not lead to further increase. Both detection methods were equally suitable for the measurement of IL 1 biological activity in purified and crude preparations. In contrast to the conventional thymocyte comitogenic assay, the fibroblasts in this assay did not proliferate in response to IL 2 or IL 6. Fibroblasts were weakly stimulated by recombinant (rec) tumor necrosis factor (rec TNF-alpha); they did, however, not proliferate in response to mitogens, lipopolysaccharide, rec granulocyte-macrophage colony stimulating factor (rec GM-CSF), macrophage-CSF, rec interferon-gamma, insulin or transferrin. The detection of IL 1 activity by crystal violet staining of human dermal fibroblasts was easier and faster than by measurem...Continue Reading

  • References6
  • Citations13

Mentioned in this Paper

Structure of Calf of Leg
Tumor Necrosis Factor-alpha
Specimen Type - Fibroblasts
Thymidine
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-1
Cattle calf (organism)
Transferrin
Thymocyte
Bos taurus

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