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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

Cell proliferation. Immunohistochemical staining of the proliferation marker Ki-67 is shown. A monoclonal mouse antibody against human Ki-67 was used to detect proliferating cells (green). A DAPI counter stain was also performed to determine the nuclei of the cells. Images were obtained at the corresponding wavelength and merged together to show the localization of Ki-67 signal within the nucleus of proliferating cells (white arrow head). At day 7, Ki-67 was strictly concentrated in the basal layer (a). At days 14, 21, and 28, the signal became more widely distributed across the whole epidermis. This distribution was associated with an epidermal loss of structure during the 4-week follow-up (day 28) (d). A control staining without using the anti Ki-67 antibody is shown in the upper right margin of each image.
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Figure 2: Cell proliferation. Immunohistochemical staining of the proliferation marker Ki-67 is shown. A monoclonal mouse antibody against human Ki-67 was used to detect proliferating cells (green). A DAPI counter stain was also performed to determine the nuclei of the cells. Images were obtained at the corresponding wavelength and merged together to show the localization of Ki-67 signal within the nucleus of proliferating cells (white arrow head). At day 7, Ki-67 was strictly concentrated in the basal layer (a). At days 14, 21, and 28, the signal became more widely distributed across the whole epidermis. This distribution was associated with an epidermal loss of structure during the 4-week follow-up (day 28) (d). A control staining without using the anti Ki-67 antibody is shown in the upper right margin of each image.

Mentions: Skin samples were cultured for a 4-week follow-up. Every week, 1 sample was removed, formalin fixed, paraffin embedded, and sliced into 4-μm sections for further analysis. A change in the number of Ki-67-positive keratinocytes was not observed, whereas a change was observed in the organization and epidermal localization. At the beginning of skin culturing, a high amount of Ki-67-positive cells were observed within the basal layer of the epidermis. During maturation of the tissue specimen, Ki-67-positive cells were more and more distributed in a randomized fashion within the complete epidermis (Fig 2).


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)

Cell proliferation. Immunohistochemical staining of the proliferation marker Ki-67 is shown. A monoclonal mouse antibody against human Ki-67 was used to detect proliferating cells (green). A DAPI counter stain was also performed to determine the nuclei of the cells. Images were obtained at the corresponding wavelength and merged together to show the localization of Ki-67 signal within the nucleus of proliferating cells (white arrow head). At day 7, Ki-67 was strictly concentrated in the basal layer (a). At days 14, 21, and 28, the signal became more widely distributed across the whole epidermis. This distribution was associated with an epidermal loss of structure during the 4-week follow-up (day 28) (d). A control staining without using the anti Ki-67 antibody is shown in the upper right margin of each image.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Cell proliferation. Immunohistochemical staining of the proliferation marker Ki-67 is shown. A monoclonal mouse antibody against human Ki-67 was used to detect proliferating cells (green). A DAPI counter stain was also performed to determine the nuclei of the cells. Images were obtained at the corresponding wavelength and merged together to show the localization of Ki-67 signal within the nucleus of proliferating cells (white arrow head). At day 7, Ki-67 was strictly concentrated in the basal layer (a). At days 14, 21, and 28, the signal became more widely distributed across the whole epidermis. This distribution was associated with an epidermal loss of structure during the 4-week follow-up (day 28) (d). A control staining without using the anti Ki-67 antibody is shown in the upper right margin of each image.
Mentions: Skin samples were cultured for a 4-week follow-up. Every week, 1 sample was removed, formalin fixed, paraffin embedded, and sliced into 4-μm sections for further analysis. A change in the number of Ki-67-positive keratinocytes was not observed, whereas a change was observed in the organization and epidermal localization. At the beginning of skin culturing, a high amount of Ki-67-positive cells were observed within the basal layer of the epidermis. During maturation of the tissue specimen, Ki-67-positive cells were more and more distributed in a randomized fashion within the complete epidermis (Fig 2).

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