Differentiation of immortalized human keratinocytes in vivo

Krueger, GG; Jorgensen, Cynthia M.; Matsunami, Norisada; Morgan, J.; Liimatta, Andrew P.; Petersen, MJ

doi:10.1016/s0923-1811(98)83815-2

Cited by 1 publication

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“…Immunohistochemical assessment of markers of differentiation (keratins, involucrin, basement membrane components), revealed no differences between grafts with immortalized and normal keratinocytes. Most importantly, there was no evidence of loss of expression of the marker transgene (lacZ) in vivo (32,33) in the immortalized cells. In contrast, it is our experience and that of other investigators that expression in nonimmortalized cells decreases in vivo and is usually absent by six weeks (34–40), even when > 99% of the transplanted cells express the transgene (34).…”

Section: Analysis Of the Loss Of Transgene Expression In Vivomentioning

confidence: 98%

Biologic Aspects of Expression of Stably Integrated Transgenes in Cells of the Skin In Vitro and In Vivo

Krueger

Morgan

Petersen

1999

Proc Assoc American Phys

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The observation that transgenes can be stably integrated into the genome of fibroblasts using recombinant retroviruses enhanced interest in using these cells as a vector for gene therapy. This enthusiasm has lessened during the past 8 years, not because skin has lost the features that make it attractive for gene therapy, but rather because stable transgene expression in vivo has not been achieved. All investigators who have used genetically modified fibroblasts to study in vivo aspects of gene therapy have shown a decrease in transgene expression with time. This contrasts with transgene expression in similarly transduced fibroblasts in vitro, where expression is not lost or is lost very slowly. We have initiated an approach to bring further understanding to the biology of transgene expression by fibroblasts carrying stably integrated transgenes in an in vivo setting. Experiments described permit the following conclusions. Expression by and survival of genetically modified fibroblasts a) requires a persistent matrix scaffold in in vivo settings; b) is prolonged if the matrix is allowed to mature in vitro; c) is enhanced if the matrix is partially sequestered behind a coating of normal fibroblasts; and d) can be substantively prolonged in vivo by immortalizing the cells. These observations support the notion that prolonged expression of transgenes by fibroblasts can be achieved in vivo and that gene therapy utilizing fibroblasts and other cells of the skin has clinical utility.

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Section: Analysis Of the Loss Of Transgene Expression In Vivomentioning

confidence: 98%