Immunocytochemical methods in cell cultures for evaluation of cell cycle and differentiation in normal and neoplastic tissues

Primary cell culture, tissue culture, and established cell line culture methods are addressed in this paper. These methods are easily applicable to the pathology laboratory. Sterile conditions to obtain, process, disaggregate the surgical specimen, and incubate the cells are emphasized. Passage to multiwell-antisera saving plates, specific streptavidin-biotin-peroxidase and phosphatase-antiphosphatase method modifications, and bromodeoxyurydine incorporation in cell culture medium are described to evaluate the cellular proliferation. We also discuss new features about conservation of easily-preserved slides that can be filed as biopsies. Satisfactory results have been obtained using immunohistochemical methods in cell cultures with antibodies against vimentin, glial fibrillary acidic protein, muscle actin, proliferating cell nuclear antigen, bromodeoxyurydine, and AE1 keratin.