Experimental Study On Biological Characteristics And Wound Grafting Of SV40 Transformed Human Keratinocytes

Characterized by low proliferating rate and limited life span in routine in vitro culture system, the growth pattern of normal human keratinocytes reflects the major disadvantages of using this cell type in the related basic research and the clinical application. In previous studies, however, many SV40 transformed human keratinocyte lines were found to have a long-term in vitro life spans, retained normal differentiation functions and non-tumorigenic feature. These transformants, with better proliferation ability and long-term life span, may have significance in tissue repair research. Based upon this consideration, a series of in vitro and in vivo experiments were conducted to investigate the feasibility of SV40 transformed keratinocytes in wound repair by means of cultured sheet grafting. The experiments are mainly classified into three parts: (1) in vitro transformation of human keratinocytes by SV40 virus, (2) identification and biological characterization of the transformants, and (3) preliminary application of transformed keratinocytes in excisional wound grafting. The results demonstrated that:I. After infection with SV40, transformed cell colonies were formed and cell lines were obtained by colony isolation and further in vitro cultivation. For several cell lines obtained, the viral genes encoding SV40 Large T antigen were integrated into the genomic DNA of the host keratinocytes determined by Southern blotting and SV40 Large T antigen was expressed in nucleus region demonstrated by indirect immunofluorescence. The transformants were...