Gingival keratinocyte grafts are usually cultured using 3T3 mouse fibroblasts as feeder cells and fetal calf serum as growth factor. These additives entail risks due to xenogenic DNA and protein. Therefore the explant and the disperse culture technique free of feeder cells were compared, and autogenous human serum was tested. Twelve halved gingival biopsies were trypsinized and cultured as single-cell suspensions, the other halves were cultured as explants. Six halved biopsies were cultured in autogenous serum, the other halves in fetal calf serum. Growth, morphology and cell biological aspects were compared. The single-cell suspensions did not form a confluent epithelial layer, whereas all explants formed confluent primary gingival keratinocyte cultures. The keratinocytes' growth in autogenous serum was equivalent to that in fetal calf serum. Morphology and cytokeratin expression were identical. The explant technique combined with autogenous serum can be used for culturing gingival autografts as well as for individual cultures for special issues.