Human keratinocyte cell culture for the burns patients--a preliminary report

One of the major problems in extensive burns is the relative lack of available donor sites for skin grafting. Keratinocyte cell culture in the laboratory was carried out successfully in Singapore General Hospital and shows promise as an alternative source for skin replacement. Our experience further proved that a fibroblast base is necessary for keratinocyte cell culture. It is observed that heterogenous cell source can grow concurrently and become confluent. The colony forming efficiency from trypsinized skin is about 1-4% in primary cultures and 35-40% in secondary cultures. The time taken to reach confluency are 20-21 days and 10-12 days respectively. The thickness of cultured skin is estimated as 0.5 mm (5-6 layers) under light microscope. The size of the harvested cultured skin is approximately one third of the cultured area due to contraction. The expansion ratio before shrinkage is estimated to be approximately 6000-fold based on our data.