Long-term transplantation of canine keratinocytes made resistant to G418 through retrovirus-mediated gene transfer.

Flowers, Mary E.D.; Stockschlaeder, Marcus; Schuening, FG; Niederwieser, Dietger; Hackman, Robert C.; Miller, A. Dusty; Storb, Rainer

doi:10.1073/pnas.87.6.2349

Cited by 58 publications

(21 citation statements)

References 28 publications

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“…Further, these cells persisted for 11 months after grafting; they were identified as male cells in a female host. Such long-term persistence of grafted male cells in females had been previously reported [53], as well as the persistence for 27 years of male cells in the blood of females who had given birth to male offspring [54]. Such data suggest that stem cells may exist in all tissues, that these stem cells may have the capacity to become any tissue [55][56][57], and that stem cells have a very long life.…”

Section: Cells Were Found In All 13-day and 60-day Adult Tissues Derimentioning

confidence: 67%

Somatic Epidermal Stem Cells Can Produce Multiple Cell Lineages During Development

2002

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It has been demonstrated that several types of somatic stem cells have the remarkable capacity to differentiate into other types of tissues. We demonstrate here that stem cells from the skin, the largest organ of the body, have the capacity to form multiple cell lineages during development. Using our recently developed sorting technique, we isolated viable homogenous populations of somatic epidermal stem and transient amplifying cells from the skin of 3-day old transgenic mice, who carried the enhanced green fluorescent protein transgene, and injected stem, TA, or unsorted basal epidermal cells into 3.5-day C57BL/6 blastocysts. Only the stem-injected blastocysts produced mice with GFP + cells in their tissues. We found GFP + cells in ectodermal, mesenchymal, and neural-crestderived tissues in E13.5 embryos, 13-day-old neonates, and 60-day-old adult mice, proving that epidermal stem cells survived the blastocyst injection and multiplied during development. Furthermore, the injected stem cells altered their epidermal phenotype and expressed the appropriate proteins for the tissues into which they developed, demonstrating that somatic epidermal stem cells have the ability to produce cells of different lineages during development. These data suggest that somatic epidermal stem cells may show a generalized plasticity expected only of embryonic stem cells and that environmental (extrinsic) factors may influence the lineage pathway for somatic stem cells. Thus, the skin could be a source of easily accessible stem cells that are able to be reprogrammed to form multiple cell lineages.

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Section: Cells Were Found In All 13-day and 60-day Adult Tissues Derimentioning

confidence: 67%

Somatic Epidermal Stem Cells Can Produce Multiple Cell Lineages During Development

2002

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“…Given their extraordinary proliferative capacity in culture (1)(2)(3), and the well-documented value of cultured keratinocytes in burn grafting operations (4), genetically manipulated skin keratinocytes have long been proposed as a potential vehicle to correct for a human deficiency in a circulating protein or factor (5,12,(45)(46)(47). In the first systemic delivery experiments attempted, Morgan et al (5) grafted cultured retroviral promoter-driven hGH-transfected keratinocytes to the skin of a nude mouse.…”

Section: Discussionmentioning

confidence: 99%

Transgenic studies with a keratin promoter-driven growth hormone transgene: Prospects for gene therapy

Wang

Zinkel

Polonsky

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

150

110

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Keratinocytes are potentially appealing vehicles for the delivery of secreted gene products because they can be transferred to human skin by the relatively simple procedure of grafting. Adult human keratinocytes can be efficiently propagated in culture with sufficient proliferative capacity to produce enough epidermis to cover the body surface of an average adult. However, the feasibility of delivering secreted proteins through skin grafting rests upon (i) the strength of the promoter in keratinocytes and (ii) the efficiency of protein transport through the basement membrane of the stratified epithelium and into the bloodstream. In this paper, we use transgenic technology to demonstrate that the activity of the human keratin 14 promoter remains high in adult skin and that keratinocyte-derived human growth hormone (hGH) can be produced, secreted, and transported to the bloodstream of mice with efficiency that is sufficient to exceed by an order of magnitude the circulating hGH concentration in growing children. Transgenic skin grafts from these adults continue to produce and secrete hGH stably, at Ϸ1͞10 physiological levels in the bloodstream of nontransgenic recipient mice. These studies underscore the utility of the keratin 14 promoter for expressing foreign transgenes in keratinocytes and demonstrate that keratinocytes can be used as effective vehicles for transporting factors to the bloodstream and for eliciting metabolic changes. These findings have important implications for considering the keratinocyte as a possible vehicle for gene therapy.

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“…Epidermal keratinocytes expressing transduced Factor IX were found to lose expression within 28 days after being grafted in one study (Gerrard et al, 1993) and within six weeks in another (Fenjves et al, 1996). Canine epidermal keratinocytes were found to lose expression of a neo r gene after subcutaneous implantation into donor animals, even though the transgene was still present (Flowers et al, 1990), and p-gal expression persisted for only 14 days in the same host (Stockschlaeder et al, 1994). It appears that, similarly to other cell types, keratinocytes carrying a transgene will survive after being grafted, but that virally encoded genes are gradually inactivated in vivo.…”

Section: (B) Regulated Gene Expression Allowing For Targeted Gene Delmentioning

confidence: 99%

“…Other reporter genes that have been introduced into keratinocytes include chloramphenicol acetyl transferase (CAT), luciferase, neuropeptide P, and SV40 T antigen. None of the selectable or marker genes thus far tested has had deleterious effects on keratinocyte growth potential or differentiation (Flowers et al, 1990;Garlick et al, 1991). Furthermore, transgenic mice expressing (3-gal in keratinocytes do not appear to undergo changes or transformation when compared with normal mouse keratinocytes (Sanes et al, 1986).…”

Section: (2) Reporter Genesmentioning

confidence: 99%

Keratinocyte Gene Transfer and Gene Therapy

Garlick

Fenjves

1996

Critical Reviews in Oral Biology & Medicine

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Gene therapy has moved beyond the pre-clinical stage to the treatment of a variety of inherited and acquired diseases. For such therapy to be successful, genes must be efficiently delivered to target cells and gene products must be expressed for prolonged periods of time without toxic effects to the host. This may be achieved by means of an in vivo strategy where genes are transferred directly into a host cell, or by means of an ex vivo approach through which cells are removed, cultured, targeted for gene delivery, and grafted back to the host. Several obstacles continue to delay safe and effective clinical application of gene therapy in a variety of target cells. The limited survival of transplanted cells, transient expression of transferred genes, and difficulties in targeting stem cells are technical issues requiring further investigation.Epidermal and oral keratinocytes are potential vehicles for gene therapy. Several features of these tissues can be utilized to achieve delivery of therapeutic gene products for local or systemic delivery. These qualities include: (1) the presence of stem cells; (2) the cell-, strata-, and site-specific regulation of keratinocyte gene expression; (3) tissue accessibility; and (4) secretory capacity. Such features can be exploited by the use of gene therapy strategies to facilitate: (1) identification, enrichment, and targeting of stem cells to ensure the continued presence of the transferred gene; (2) high-level and persistent transgene expression using keratinocyte-specific promoters; (3) tissue access needed for culture and grafting for ex vivo therapy and direct in vivo gene transfer; (4) secretion of transgene product for local or systemic delivery; and (5) monitoring of genetically modified tissue and removal if treatment termination is required. Optimal gene therapy strategies are being tested in a variety of tissues to treat dominant and recessive genetic disorders as well as acquired diseases such as neoplasia and infectious disease. This experience provides a basis for the application of such clinical studies to a spectrum of diseases effecting epidermal and oral keratinocytes. Gene therapy is in an early stage yet holds great promise for its ultimate clinical application.

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Long-term transplantation of canine keratinocytes made resistant to G418 through retrovirus-mediated gene transfer.

Cited by 58 publications

References 28 publications

Somatic Epidermal Stem Cells Can Produce Multiple Cell Lineages During Development

Somatic Epidermal Stem Cells Can Produce Multiple Cell Lineages During Development

Transgenic studies with a keratin promoter-driven growth hormone transgene: Prospects for gene therapy

Keratinocyte Gene Transfer and Gene Therapy

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