Development of an ex vivo human skin model for intradermal vaccination: tissue viability and Langerhans cell behaviour.
The presence of resident Langerhans cells (LCs) in the epidermis makes the skin an attractive target for DNA vaccination. However, reliable animal models for cutaneous vaccination studies are limited. We demonstrate an ex vivo human skin model for cutaneous DNA vaccination which can potentially bridge the gap between pre-clinical in vivo animal models and clinical studies. Cutaneous transgene expression was utilised to demonstrate epidermal tissue viability in culture. LC response to the culture environment was monitored by immunohistochemistry. Full-thickness and split-thickness skin remained genetically viable in culture for at least 72 h in both phosphate-buffered saline (PBS) and full organ culture medium (OCM). The epidermis of explants cultured in OCM remained morphologically intact throughout the culture duration. LCs in full-thickness skin exhibited a delayed response (reduction in cell number and increase in cell size) to the culture conditions compared with split-thickness skin, whose response was immediate. In conclusion, excised human skin can be cultured for a minimum of 72 h for analysis of gene expression and immune cell activation. However, the use of split-thickness skin for vaccine formulation studies may not be appropriate because of the nature of the activation. Full-thickness skin explants are a more suitable model to assess cutaneous vaccination ex vivo.
Ng
Keng Wooi
KW
Gene Delivery Research Group, Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3NB, UK.
Pearton
Marc
M
Coulman
Sion
S
Anstey
Alexander
A
Gateley
Christopher
C
Morrissey
Anthony
A
Allender
Christopher
C
Birchall
James
J
D000328:Adult; D000368:Aged; D000078404:Epidermal Cells; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D007271:Injections, Intradermal; D007801:Langerhans Cells / Q000166:cytology* / Q000276:immunology; D008875:Middle Aged; D009924:Organ Culture Techniques; D010957:Plasmids; D012867:Skin / Q000166:cytology* / Q000276:immunology; D014022:Tissue Survival; D019076:Transgenes; D014611:Vaccination
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