Effect of donor age on cultivation of human oral mucosal keratinocytes

Cultured mucosal grafts (CMG) are among recently developed biological grafting materials to cover large oral mucosal defects following resection of mucosal pathology. This study evaluates the effect of donor's age on the cultivation process of oral mucosal keratinocytes for grafting. Human mucosal epithelial cells were utilized and classified into three donor age groups: 3-30 years (14 patients); 31-60 years (9 patients); and >60 years (6 patients) (11 males and 18 females). Isolation and cultivation of oral mucosal keratinocytes were according to Rheinwald and Green [Cell 6 (1975) 331], originally developed for epidermal keratinocytes. Isolated primary cell lines were seeded and cultivated. Propagation of cell lines ("passages"), time period required to reach confluence, yield of cells and plating efficiency were recorded. All cells propagated well up to the fourth passage. Thereafter, a decline was observed and was more distinct with age. Period to confluence was longer among the old age group. Yield of cells in fourth passage was high among the young age group and decreased with age. Plating efficiency in passages 4-6 decreased with age. These results suggest that age-related changes in cultivation of oral keratinocytes are not general phenomena, but rather limited to the donor age of 60 years and above. In this age group all the parameters studied were adversely affected. Oral mucosal keratinocytes may be a useful model for oral aging.