Chapters authored
Optimization of the Self-Assembly Method for the Production of Psoriatic Skin Substitutes
Tissue engineering of the skin is used for various applications. However, to develop treatments for skin pathologies such as psoriasis, robust pathological skin models are needed. The purpose of the work presented in this chapter was to optimize the production of more reproducible psoriatic skin substitutes by modifying the original self-assembly method. Substitutes were produced according to the self-assembly method partially modified. The culture flasks of 25 cm2 were replaced by 6-well and 12-well plates. Fibroblasts were cultured in 6-well and 12-well plates with ascorbic acid until they form manipulable sheets, which were superimposed and incubated for 7 days to form a dermal layer. Afterwards, keratinocytes were seeded on the dermal layer forming an epidermal layer. Then, the substitutes were raised to the air-liquid interface and cultured 21 days before being analyzed. Analyses demonstrated that psoriatic substitutes have a significantly thicker epidermis than healthy substitutes and the persistence of nuclear structures in corneocytes, with original and both modified methods. Immunofluorescence markers such as filaggrin, loricrin, and keratin 14 have confirmed these results. However, some differences were observed in substitutes produced with 12-well plates. Modifications made to the original method for the production of psoriatic substitutes are effective and lead to highly reproducible substitutes more suitable for pharmacological testing.
Part of the book: Cell Culture