Background: Co-culture techniques have important role in the study of cellular interactions. The ability of distinguishng co-cultured cells facilitates evaluation of these cultures. P388D1 cells are macrophage like cells but morphologic appearance of these cells is like lymphoblasts. In evaluation of P388D1 and bone marrow cells co-culture, Different antigens on P388D1 cells surface could help to distinguish these cells from bone marrow cells.

Materials and Methods : In order to prepare the antibody against P388D1 cells, rabbits were immunized by intraperitoneal and intravenous injections of these cells. After getting blood, serum immunoglobulins were precipitated in ammonium sulfate solution. Before and after each precipitation, serum protein electrophoresis was performed and protein was assayed by Bradford method. Antibodies, which had a cross reactivity with bone marrow cells were removed by several absorptions with bone marrow cells. The presence of antibodies on cell surface was detected by indirect immunoflurescence labeling technique. Complement mediated cytotoxicity of antibodies was also assayed.

Results: The results indicate that this antibody solution has no detectable cross reaction with bone marrow cells, using indirect immunflurescence labeling technique, while it has positive reaction against P388D1 cells.

Conclusion: By means of these antibodies we would be able to recognize or remove P388D1 cells from bone marrow cell population.