A new system for the cultivation of keratinocytes on acellular human dermis with the use of fibrin glue and 3T3 feeder cells

The growth of human keratinocytes on human acellular dermis in 4 different culture systems was compared. Epidermis was completely separated and remove d from dermis after skin samples had been soaked in 0.1% trypsin at 4 degre es C for 1 week. Forty pieces of saline washed dermis, I cm(2) each, mere r andomized into 4 groups: in group A, human keratinocytes that had undergone 2 to 3 cell passages were seeded (30 x 10(4) cell/cm(2)) onto the dermis a nd sprayed with a thin layer of fibrin glue and proliferative 3T3 feeder ce lls that had been growing separately on the culture dish; in group B, the d ermis was only sprayed with fibrin glue; in group C, the dermis was treated with 3T3 cells only; and in group D, the dermis was not sprayed with anyth ing. The dermis samples in all groups were raised on a grid to provide an a ir-liquid culture system. Histology results of the composite grafts at 2 we eks were assessed as having either scanty colonies of keratinocytes (SCK), continuous stratified epithelium (CSE), or no observable keratinocyte growt h. Eight out of the ten dermis samples (80%) in group A demonstrated CSE, a nd 30% of the samples in group B showed SCK There were 10% SCK and 20% CSE in group C, and in group D, 30% SCK and 10% CSE were found. The good result s in group A indicated that the fibrin glue facilitated the seeding efficie ncy of the keratinocytes on the dermis and that the vital factors released from the 3T3 feeder cells enhanced the growth and differentiation of the ke ratinocytes. This model provides an optimal system for the cultivation of k eratinocytes on acellular dermis.