title: Evolutive skin reconstructions: from the dermal collagen-glycosaminoglycan-chitosane substrate to an immunocompetent reconstructed skin.
abstract: The development of human skin models that have the same properties as genuine human skin is of particular significance. Very promising skin models are the three-dimensional artificial skin constructs, which, similar to genuine skin, consist of an epidermis of differentiated keratinocytes and a dermis. A skin equivalent based on a collagen-glycosaminoglycan-chitosan dermal substrate has been developed to meet the growing demand in tissue engineered skin equivalents. We used this model to investigate whether CD34-generated Langerhans/dendritic cell precursors could be integrated into this skin equivalent model and pursue their differentiation without addition of cytokine and growth factor. To address the issue of dendritic cell (DC) differentiation, an endothelialized skin equivalent coculture model was used to study the behaviour of haematopoietic progenitor cells (HPC) in epidermal and dermal environments. CD34(+) HPC were cultured for 6 days with GM-CSF, TGFbeta1 and TNFalpha and seeded in the endothelialized skin equivalent at different time points to favour dermal or epidermal integration. This integration (after keratinocyte seeding, only and in absence of exogenous GM-CSF, TNFalpha, TGFbeta1) gave rise both cutaneous DC, i.e. epidermal Langerhans cells (CD1a(+), HLA-DR(+)) and dermal DC (DC-SIGN(+), HLA-DR(+)) while endothelial cells are sufficiently activated to acquire HLA-DR expression. For the first time, the presence of a living dermal equivalent could provide a more complex environment integrating vascular components to study the differentiation of interstitial DC in a dermis equivalent. Such sophisticated skin equivalent may clarify some intriguing aspects of the numerous regulatory mechanisms controlling skin homeostasis.
authors.affiliation: Inserm ex-U 346/ EA no 37-32 UCBL1, Hôpital Edouard Herriot, Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Dezutter-Dambuyant
authors.forename: Annie
authors.initials: A
authors.lastname: Black
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bechetoille
authors.forename: Charbel
authors.initials: C
authors.lastname: Bouez
authors.forename: Sylvie
authors.initials: S
authors.lastname: Maréchal
authors.forename: Céline
authors.initials: C
authors.lastname: Auxenfans
authors.forename: Valérie
authors.initials: V
authors.lastname: Cenizo
authors.forename: Pascale
authors.initials: P
authors.lastname: Pascal
authors.forename: Eric
authors.initials: E
authors.lastname: Perrier
authors.forename: Odile
authors.initials: O
authors.lastname: Damour
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D002417:Cattle; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D018920:Coculture Techniques; D003094:Collagen / Q000737:chemistry*; D003713:Dendritic Cells / Q000166:cytology; D020405:Dermis / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry*; D015603:Keratinocytes / Q000166:cytology; D012867:Skin / Q000473:pathology*
chemical_list: D018952:Antigens, CD34; D006025:Glycosaminoglycans; D003094:Collagen; D048271:Chitosan