A new method for studying epidermalization in vitro.
A new method for studying epidermalization in vitro is described. It consists of inserting a punch biopsy that serves as a source of epidermis into dermal equivalent freshly made up, with fibroblasts mixed in a collagen matrix. Fibroblasts cling to collagen fibrils and contract the matrix, leading in 3 days to a resistant dermal equivalent holding the punch biopsy firmly in place. At day 5, a culture medium favouring epidermal growth was used and a fringe of a new epidermis appeared around the punch, the area of which grew linearly with time. This new epidermis showed a pattern of differentiation similar to epidermis in vivo, with cuboidal basal cells, keratohyalin granules, membrane coating granules and the expression of the 65-67 kd keratin subset. The method seems to combine the advantages of the explant technique and of classical keratinocyte cultures, providing the researcher with a large quantity of differentiated epidermis, the pharmacologist with simple and quantitative system in which to study modifications of growth and differentiation of epidermis, and the plastic surgeon with a possible material for skin grafting.
B
B
Coulomb
P
P
Saiag
E
E
Bell
F
F
Breitburd
C
C
Lebreton
M
M
Heslan
L
L
Dubertret
D001706:Biopsy; D002455:Cell Division; D046508:Culture Techniques / Q000379:methods; D000078404:Epidermal Cells; D004817:Epidermis / Q000032:analysis / Q000648:ultrastructure; D006801:Humans; D007633:Keratins / Q000032:analysis; D008854:Microscopy, Electron
D007633:Keratins