Background: Our previous studies revealed presence of a number of growth
factors and cytokines in human skin tissue fluid/lymph (TF/L) at levels higher than in serum. This prompted us to study whether TF/L may have a regulatory effect on keratinocyte (KC) growth. Aim: To study the effect of TF/L on proliferation and differentiation of KC and their stem cell markers expression.
Material/Methods: KC were isolated from lower limb skin and were cultured
for 1 to 7 days in TF/L. Phenotypes were identifi ed using antibodies against
p63, CD29, Ki67 and PCNA. Blocking of cytokines with antibodies helped to
estimate which cytokine stimulated KC proliferation and differentiation.
Results: KC cultured in TF/L showed higher than in controls percentage of
dividing and elongated cells from basal layer as well as lower percentage of
differentiated cells from upper layers. Higher percentage of p63 and CD29
positive cells was also observed. Neutralization of IL-1b, IL-6, TNF-a, KGF caused decrease in percentage of mitotic cells. Neutralization of KGF decreased percentage of p63 and CD29 positive cells.
Conclusions: TF/L cytokines have a stimulating effect on proliferation of basal
KC but not on their differentiation. KGF turned to be a strong stimulator.

Keywords: biostatic grafts, composite tissue transplantation, Limb Transplantation