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A human full-skin culture system for interventional studies.

Steinstraesser L, Rittig A, Gevers K, Sorkin M, Hirsch T, Kesting M, Sand M, Al-Benna S, Langer S, Steinau HU, Jacobsen F - Eplasty (2009)

Bottom Line: The aim of this study was to develop an effective surrogate model in which ex vivo full-thickness organ culture experiments may be performed.Transgene expression was demonstrated to be time dependent.This model chamber presents a convenient, easy-to-use, and robust model in which ex vivo full-thickness organ culture experiments may be performed.

View Article: PubMed Central - PubMed

Affiliation: Department for Plastic Surgery, Burn Center, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum, Germany.

ABSTRACT

Objective: Novel approaches to bridge the gap between clinical studies and experimental basic research of skin physiology are urgently needed. The aim of this study was to develop an effective surrogate model in which ex vivo full-thickness organ culture experiments may be performed.

Methods: Human full skin from patients was placed into a stainless steel chamber and cultured at an air-liquid interphase for 4 weeks. Samples were evaluated every week by HE-staining and immunohistochemical characterization. Epidermal gene transfer kinetics was performed as an interventional study.

Results: This ex vivo chamber model maintained the physiologic and histologic properties of the skin explants for 4 weeks. This indicated the model's acceptable ex vivo physiologic validity. No epidermolysis was observed, and both basal lamina and blood vessels were detected within all tissue samples. Transgene expression was demonstrated to be time dependent.

Conclusion: This model chamber presents a convenient, easy-to-use, and robust model in which ex vivo full-thickness organ culture experiments may be performed.

No MeSH data available.


Related in: MedlinePlus

Skin culture model. The images of the upper row show the stainless steel chamber model from the epidermal (a) and the dermal (b) side of view. The macroscopic appearance of the epidermal wounding procedure is also displayed (a). After 4 weeks of culture and partial depletion of the epidermal layer, skin specimens have been topically transduced by using 108 IU of LacZ-adenoviral vectors. Samples were taken further 2 days later and transgene expression was visualized by using X-gal staining. Only tissue specimen with removed epidermis showed transgene expression after the topical application of the virus (d, blue-stained cells), indicating intact skin barrier only for tissue without epidermolysis (c). Images are at 100-fold magnification; scale bar displays 100 μm.
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Figure 6: Skin culture model. The images of the upper row show the stainless steel chamber model from the epidermal (a) and the dermal (b) side of view. The macroscopic appearance of the epidermal wounding procedure is also displayed (a). After 4 weeks of culture and partial depletion of the epidermal layer, skin specimens have been topically transduced by using 108 IU of LacZ-adenoviral vectors. Samples were taken further 2 days later and transgene expression was visualized by using X-gal staining. Only tissue specimen with removed epidermis showed transgene expression after the topical application of the virus (d, blue-stained cells), indicating intact skin barrier only for tissue without epidermolysis (c). Images are at 100-fold magnification; scale bar displays 100 μm.

Mentions: Skin explants from healthy patients were prepared and cultured for at least 4 weeks. For epidermal wounding, the epidermis was scored by using a 6-mm punching knife and a scalpel. Fifty microliters of a 0.2% dispase solution was dropped onto the scored epidermis. After 3 hours at 37°C, the epidermis within the 6-mm area was completely removed (Fig 6a). Adenoviral vectors (108 IU) were topically applied with or without the removal of the epidermis. Two days later, tissue biopsy specimens were harvested. Following this, 10-μm tissue sections were taken and X-gal staining was performed.


A human full-skin culture system for interventional studies.

Steinstraesser L, Rittig A, Gevers K, Sorkin M, Hirsch T, Kesting M, Sand M, Al-Benna S, Langer S, Steinau HU, Jacobsen F - Eplasty (2009)

Skin culture model. The images of the upper row show the stainless steel chamber model from the epidermal (a) and the dermal (b) side of view. The macroscopic appearance of the epidermal wounding procedure is also displayed (a). After 4 weeks of culture and partial depletion of the epidermal layer, skin specimens have been topically transduced by using 108 IU of LacZ-adenoviral vectors. Samples were taken further 2 days later and transgene expression was visualized by using X-gal staining. Only tissue specimen with removed epidermis showed transgene expression after the topical application of the virus (d, blue-stained cells), indicating intact skin barrier only for tissue without epidermolysis (c). Images are at 100-fold magnification; scale bar displays 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2627306&req=5

Figure 6: Skin culture model. The images of the upper row show the stainless steel chamber model from the epidermal (a) and the dermal (b) side of view. The macroscopic appearance of the epidermal wounding procedure is also displayed (a). After 4 weeks of culture and partial depletion of the epidermal layer, skin specimens have been topically transduced by using 108 IU of LacZ-adenoviral vectors. Samples were taken further 2 days later and transgene expression was visualized by using X-gal staining. Only tissue specimen with removed epidermis showed transgene expression after the topical application of the virus (d, blue-stained cells), indicating intact skin barrier only for tissue without epidermolysis (c). Images are at 100-fold magnification; scale bar displays 100 μm.
Mentions: Skin explants from healthy patients were prepared and cultured for at least 4 weeks. For epidermal wounding, the epidermis was scored by using a 6-mm punching knife and a scalpel. Fifty microliters of a 0.2% dispase solution was dropped onto the scored epidermis. After 3 hours at 37°C, the epidermis within the 6-mm area was completely removed (Fig 6a). Adenoviral vectors (108 IU) were topically applied with or without the removal of the epidermis. Two days later, tissue biopsy specimens were harvested. Following this, 10-μm tissue sections were taken and X-gal staining was performed.

Bottom Line: The aim of this study was to develop an effective surrogate model in which ex vivo full-thickness organ culture experiments may be performed.Transgene expression was demonstrated to be time dependent.This model chamber presents a convenient, easy-to-use, and robust model in which ex vivo full-thickness organ culture experiments may be performed.

View Article: PubMed Central - PubMed

Affiliation: Department for Plastic Surgery, Burn Center, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum, Germany.

ABSTRACT

Objective: Novel approaches to bridge the gap between clinical studies and experimental basic research of skin physiology are urgently needed. The aim of this study was to develop an effective surrogate model in which ex vivo full-thickness organ culture experiments may be performed.

Methods: Human full skin from patients was placed into a stainless steel chamber and cultured at an air-liquid interphase for 4 weeks. Samples were evaluated every week by HE-staining and immunohistochemical characterization. Epidermal gene transfer kinetics was performed as an interventional study.

Results: This ex vivo chamber model maintained the physiologic and histologic properties of the skin explants for 4 weeks. This indicated the model's acceptable ex vivo physiologic validity. No epidermolysis was observed, and both basal lamina and blood vessels were detected within all tissue samples. Transgene expression was demonstrated to be time dependent.

Conclusion: This model chamber presents a convenient, easy-to-use, and robust model in which ex vivo full-thickness organ culture experiments may be performed.

No MeSH data available.


Related in: MedlinePlus