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Genetic modification of cultured skin substitutes by transduction of human keratinocytes and fibroblasts with platelet‐derived growth factor‐A
Author(s) -
Supp Dorothy M,
Bell Sheila M,
Morgan Jeffrey R,
Boyce Steven T
Publication year - 2000
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1046/j.1524-475x.2000.00026.x
Subject(s) - wound healing , keratinocyte , growth factor , transduction (biophysics) , genetic enhancement , human skin , transforming growth factor , skin grafting , glycosaminoglycan , fibroblast , skin repair , microbiology and biotechnology , chemistry , platelet derived growth factor , cell culture , cancer research , immunology , biology , medicine , gene , platelet derived growth factor receptor , biochemistry , receptor , surgery , genetics
Gene therapy promises the potential for improved treatment of cutaneous wounds. This study evaluated whether genetically modified cultured skin substitutes can act as vehicles for gene therapy in an athymic mouse model of wound healing. Human keratinocytes and fibroblasts were genetically engineered by retroviral transduction to overexpress human platelet‐derived growth factor‐A chain. Three types of skin substitutes were prepared from collagen‐glycosaminoglycan substrates populated with fibroblasts and keratinocytes: HF–/HK–, containing both unmodified fibroblasts and keratinocytes; HF–/HK+, containing unmodified fibroblasts and modified keratinocytes; and HF+/HK–, containing modified fibroblasts and unmodified keratinocytes. Skin substitutes were cultured for two weeks before grafting to full‐thickness wounds on athymic mice. The modified skin substitutes secreted significantly elevated levels of platelet‐derived growth factor throughout the culture period. Expression of retroviral platelet‐derived growth factor‐A mRNA was maintained after grafting to mice, and was detected in all HF–/HK+ grafts and one HF+/HK– graft at two weeks after surgery. Although no differences were seen between control and modified grafts, the results suggest that genetically modified cultured skin substitutes can be a feasible mechanism for cutaneous gene therapy. The cultured skin model used for these studies has advantages over other skin analogs containing only epidermal cells; because it contains both fibroblasts and keratinocytes, it therefore offers greater opportunities for genetic modification and potential modulation of wound healing.