Biological and physical factors influencing the successful engraftment of a cultured human skin substitute

Parenteau, Nancy L.; Sabolinski, Michael L.; Prosky, Stefan; Nolte, Cynthia J. M.; Oleson, M. A.; Kriwet, Katrin; Bilbo, Patrick

doi:10.1002/(sici)1097-0290(19961005)52:1<3::aid-bit1>3.0.co;2-p

Cited by 35 publications

(10 citation statements)

References 37 publications

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“…During topical selection, the migration and proliferation of other skin cells, such as fibroblasts and endothelial cells, which are important for successful grafting, might be impeded by topical colchicine application. If this is a problem in vivo, composite rafts com posed of various skin cells (i.e., both MDR-transduced keratinocytes and fibroblasts) could be generated for grafting (Parenteau et al, 1996). Furtherm ore, the appropriate colchicine dose for topical selection of MDR-transduced skin grafts must be determined by titration studies.…”

Section: Discussionmentioning

confidence: 99%

Selection of Keratinocytes Transduced with the Multidrug Resistance Gene in an in Vitro Skin Model Presents a Strategy for Enhancing Gene Expression in Vivo

Pfützner

Hengge²,

Joari³

et al. 1999

Human Gene Therapy

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In gene therapy studies, achieving prolonged, high-level gene expression in a significant percentage of cells has been difficult. One solution to enhance expression would be to select for cells expressing both the desired gene and a linked selectable marker gene in a bicistronic vector. As a potential target tissue, the skin is easily accessible for safe topical application of a selecting agent that could lead to significant gene expression in a high percentage of keratinocytes. To test the feasibility of such an approach, a skin raft culture model was developed. Human keratinocytes were transduced with the multidrug resistance (MDR) gene, which confers resistance to a variety of cytostatic and antimitotic compounds, such as colchicine. While growing on acellular dermis, transduced keratinocytes were treated with various doses of colchicine (10-50 ng/ml). Colchicine treatment increased the percentage of keratinocytes expressing MDR to almost 100% in raft cultures, Significantly, keratinocytes in colchicine-treated, MDR-transduced raft cultures were able to proliferate normally and form a stratified, differentiated epidermis. This model suggests that topical selection for MDR-expressing keratinocytes in vivo should be feasible without hampering the biologic integrity of skin. Thus, topical selection leading to enhanced expression of a desired gene, linked to a resistance gene, holds future promise for skin gene therapy.

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Section: Discussionmentioning

confidence: 99%

Selection of Keratinocytes Transduced with the Multidrug Resistance Gene in an in Vitro Skin Model Presents a Strategy for Enhancing Gene Expression in Vivo

Pfützner

Hengge²,

Joari³

et al. 1999

Human Gene Therapy

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show abstract

“…Over the past three decades, composites of cultured cells and biomaterials have been investigated for potential use as tissue engineered skin substitutes1–3 Regeneration of skin tissues has been examined with a variety of dermal analogs including collagen-glycosaminoglycan sponges,4,5 collagen gels,6,7 hyaluronic acid derivatives,8 and synthetic polymers 9. Although these composite scaffolds have demonstrated efficacious results in clinical applications, problems with mechanical stability, prolonged healing times, scarring, and limited tissue functionality persist 10.…”

Section: Introductionmentioning

confidence: 99%

Designing tailored biomaterial surfaces to direct keratinocyte morphology, attachment, and differentiation

Bush

Driscoll

Soto

et al. 2008

J Biomedical Materials Res

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Precisely engineering the surface chemistry of biomaterials to modulate the adsorption and functionality of biochemical signaling molecules that direct cellular functions is critical in the development of tissue engineered scaffolds. Specifically, this study describes the use of functionalized self-assembled monolayers (SAMs) as a model system to assess the effects of biomaterial surface properties on controlling fibronectin (FN) conformation and concentration as well as keratinocyte function. By systematically analyzing FN adsorption at low and saturated surface densities, we distinguished between SAM dependent effects of FN concentration and conformation on presenting cellular binding domains that direct cellular functions. Quantitative image analyses of immunostained samples showed that modulating the availability of the FN synergy site directly correlated with changes in keratinocyte attachment, spreading, and differentiation, through integrin mediated signaling mechanisms. The results of this study will be used to elucidate design features that can be incorporated into dermal equivalents and percutaneous implants to enhance the rate of reepithelialization and tissue regeneration. Furthermore, these findings indicate that SAM based model systems are a valuable tool for designing and investigating the development of scaffolds that regulate the conformation of extracellular matrix cues and cellular functions that accelerate the rate of tissue regeneration.

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“…This problem was also observed when we used a similar strategy with mouse growth hormone (mGH), in an attempt to have a more homologous model [Peroni et al, in preparation]. However, this in vitro loss of secretion could be completely circumvented by substituting a conventional grafting methodology, based on the classical study of Barrandon et al [1988], with an organotypic raft culture model [Garlick and Taichman, 1994;Parenteau et al, 1996;Kolodka et al, 1998]. In addition, we obtained much higher in vitro secretion levels (up to 11 µg mGH/10 6 cells/day).…”

Section: Keratinocytesmentioning

confidence: 99%

Animal Models for Growth Hormone Gene Therapy

Peroni¹,

Gout²,

Bartolini

2005

CGT

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Treatment of growth hormone (GH) deficiency via parenteral administration of recombinant hGH has greatly benefited from recombinant DNA technology allowing production of practically unlimited amounts of the pure hormone. However, an alternative approach that may lead to correction of the clinical defect is presented by hGH gene transfer into somatic cells of the patient, either ex vivo or in vivo. This approach has not only the potential advantage of circumventing repetitive injections of the hormone and its laborious isolation and purification processes, but can also, in principle, provide a mechanism of hormone delivery that resembles the natural process. GH gene therapy has not reached the clinics yet, but several interesting and promising animal models for this treatment have been developed and studied. They are not only potentially useful for elucidation of the still unresolved mechanism of sustained in vivo gene product delivery, but also for opening the way to therapy of other protein deficiencies for which gene therapy may be the only viable option. This review article describes, analyzes and compares the major animal models of GH gene therapy that have been developed in the last two decades.

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Biological and physical factors influencing the successful engraftment of a cultured human skin substitute

Cited by 35 publications

References 37 publications

Selection of Keratinocytes Transduced with the Multidrug Resistance Gene in an in Vitro Skin Model Presents a Strategy for Enhancing Gene Expression in Vivo

Selection of Keratinocytes Transduced with the Multidrug Resistance Gene in an in Vitro Skin Model Presents a Strategy for Enhancing Gene Expression in Vivo

Designing tailored biomaterial surfaces to direct keratinocyte morphology, attachment, and differentiation

Animal Models for Growth Hormone Gene Therapy

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