Six cell surface glycoproteins defined by monoclonal antibodies were selected for study on human epidermal cells. In tests on tissue sections, three of the glycoproteins [J143 (gp140/30); T43 (gp85/36); H99 (gp38)] were expressed in the basal cell layer of the epidermis, whereas the other three glycoproteins [T179 (gp140/95); T16 (gp40/50); BT15 (gp80)] were preferentially expressed in maturing keratinocytes above the basal layer. We compared synthesis of these glycoproteins in fresh epidermis and in primary epidermal short term cultures using [35S]methionine for metabolic labeling. Synthesis of J143 was 8- to 20-fold higher and synthesis of T43 was 4- to 10-fold lower in cultured cells compared with fresh epidermis. BT15, an antigen strongly expressed on terminally differentiating keratinocytes, was synthesized at 5- to 15-fold higher levels in fresh epidermis than in cultured cells. Biosynthesis levels of H99, T179, and T16 did not change in cultured epidermal cells. Based on our findings, we propose a model of surface antigenic changes that occur during keratinocyte differentiation in vivo.