Abstract
The present study attempted to utilize the hybrid-type artificial skin (hybrid skin) as an effective transgene carrier for somatic gene therapy. We have elaborated a proinsulin-secreting hybrid skin that can be used to treat diabetic animals. Rat skin fibroblasts (RSFs) were transfected with human proinsulin cDNA-containing plasmid vectors, pBMG-Neo-Ins.1 The fibroblasts were then maintained for 2 weeks in the culture medium containing G-418 to select transgenic cells (RSF308 cells). Fibroblasts transfected with pBMG-Neo without the insert cDNA were also prepared and used as control cells (RSF1518 cells). An immuno-reactive insulin (IRI) level in the culture medium of RSF308 cells was 319 ± 22.8 µIU/106 cells/h (mean ± SD). A hybrid skin was reconstituted in a 35-mm plastic dish by incorporating 2 x 106 rat fibroblasts into 2 ml of three-dimensional lattices of type I collagen (collagen gel) and then placing 106 rat keratinocytes on the gel. We made three types of hybrid skin: (a) a hybrid skin prepared w...
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