Method : In vitro reconstitution of skin equivalent
of epithelial proliferation
of epithelial differentiation
Example : evaluation of a product or ray meant to have healing properties
Implementation of a dermal equivalent by mixing type 1 collagen with fibroblasts : formation of a lattice by collagen condensation in a sterilizer/drying oven, cupboard/incubator at 37°C.
Epidermisation of equivalents by the implantation of a epidermis fragment
EPIDERMISATION
Epidermisation of equivalents by the implantation of a epidermis fragment
Change-out of culture medium two times per week for two weeks, while utilising different factors (EGF, hydrocortisone) to create favourable conditions for epidermal growth
Study of the differentiation of keratinocytes on reconstituted skin in vitro:
- Inclusion on paraffin wax for standard histological studies
- Freezing to show cytokeratins by immunochemistry;
- Fixation for ultrastructural study with electron microscope
Display of results :
- Photographs using optical and electron microscopes
- Establishment of semi-quantitative scores for cytokeratin immunochemistry
Immunohistochemical analysis of a neo-formed epithelium
Epidermisation of equivalents by the implantation of a epidermis fragment
Display of results. Establishment of semi-quantitative scores for cytokeratin immunochemistry
