A rapid method for detection of cellular proliferation using carboxyfluorescein. Assay of growth factors (IL-2, IL-1) and growth inhibiting antibodies.
The uptake of carboxyfluorescein diacetate (CFDA) into live cells was used as the basis for a simple, rapid and fully automated micromethod for determination of cell growth. The aim of the investigation was to adapt the CFDA method for detection of cell growth factors from cell culture supernatants. Thus, the biological activities of the growth factors IL-2 or IL-1 could be detected and quantitated at the same level of sensitivity as with the conventional [3H]thymidine incorporation. Similarly, the method was well suited to assay inhibition of IL-2-dependent lymphocyte growth by the monoclonal antibody anti-Tac, binding to the human lymphocyte membrane receptor for IL-2. The CFDA method proved to be rapid, reliable and well suited for several applications involving limiting numbers of cells and biological reagents.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology*; D002455:Cell Division; D005452:Fluoresceins; D006801:Humans; D007375:Interleukin-1 / Q000032:analysis*; D007376:Interleukin-2 / Q000032:analysis*; D008213:Lymphocyte Activation; D011971:Receptors, Immunologic / Q000276:immunology*; D015375:Receptors, Interleukin-2; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005452:Fluoresceins; D007375:Interleukin-1; D007376:Interleukin-2; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; C027780:carboxyfluoresceindiacetate