To use cultured mucosa as a graft of full thickness, our laboratory has been involved in the development of techniques to grow epidermis together with connective tissue. Human oral mucosa was obtained at dental surgery. Under sterile conditions
the
tissues were cut into explants of 0.1 ㎠ which were placed in the center of 24 well tissue culture dishes and incubated in a growth medium. The growth medium used for epithelial cells was MEM (Minimum Essential Medium) supplemented with 10% fetal
calf
serum, 0.5% dimethyl sulfoxide, glutamine (0.292 g/l), epidermal growth factor (40 ug/ml), cholera toxin (30 ng/ml), hydrocortisone (2 ug/ml), insulin (40 ug/ml) and transferin (5 ug/ml). The medium used for stratification of epithelial cells was
MEM
supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide and glutamine (0.292 g/l). The medium used for fibroblasts was MEM supplemented with 10% fetal calf serum. With the three types of media used alternatively, a mucosa composed of
epidermis
and connective tissue was obtained after 3 weeks of culture.
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