Coulomb B, Saiag P, Bell E, Breitburd F, Lebreton C, Heslan M, Dubertret L
Br J Dermatol. 1986 Jan;114(1):91-101. doi: 10.1111/j.1365-2133.1986.tb02783.x.
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.
本文描述了一种体外研究表皮形成的新方法。该方法包括将作为表皮来源的冲孔活检组织插入新制备的含成纤维细胞的真皮替代物中，成纤维细胞混入胶原基质。成纤维细胞附着于胶原纤维并使基质收缩，3天后形成坚韧的真皮替代物，将冲孔活检组织牢固固定在位。在第5天，使用有利于表皮生长的培养基，冲孔周围出现新表皮边缘，其面积随时间呈线性增长。这种新表皮显示出与体内表皮相似的分化模式，有立方形基底细胞、透明角质颗粒、膜被颗粒以及65 - 67kd角蛋白亚群的表达。该方法似乎结合了外植体技术和经典角质形成细胞培养的优点，为研究人员提供大量分化的表皮，为药理学家提供研究表皮生长和分化变化的简单定量系统，为整形外科医生提供一种可能用于皮肤移植的材料。
