De Vuyst Evelyne, Charlier Céline, Giltaire Séverine, De Glas Valérie, de Rouvroit Catherine Lambert, Poumay Yves
Cell and Tissue Laboratory, URPHYM, Namur Research Institute for Life Sciences, University of Namur, Namur, Belgium.
Methods Mol Biol. 2014;1195:191-201. doi: 10.1007/7651_2013_40.
This chapter provides methods suitable for the culture of primary human keratinocytes in serum-free culture conditions, starting from very small skin biopsies. It also explains procedures required for reconstruction of a stratified epidermis on polycarbonate filter, starting from keratinocytes cultured in serum-free conditions. Tissues reconstructed according to this method have been proven suitable for characterization of epidermal morphogenesis and for in vitro studies of the epidermal barrier. Utilization of the same method for successful isolation of keratinocytes from a patient suffering from Darier's disease and the reconstruction of a pathological epidermis which displays the same histological features as in vivo are also presented.
本章提供了适用于在无血清培养条件下从非常小的皮肤活检样本开始原代人角质形成细胞培养的方法。它还解释了从在无血清条件下培养的角质形成细胞开始在聚碳酸酯滤膜上重建分层表皮所需的程序。根据该方法重建的组织已被证明适用于表皮形态发生的表征和表皮屏障的体外研究。还介绍了利用相同方法成功从 Darier 病患者中分离角质形成细胞并重建具有与体内相同组织学特征的病理性表皮。
