IMPROVED BARRIER STRUCTURE FORMATION IN AIR-EXPOSED HUMAN KERATINOCYTE CULTURE SYSTEMS
IMPROVED BARRIER STRUCTURE FORMATION IN AIR-EXPOSED HUMAN KERATINOCYTE CULTURE SYSTEMS
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DOI：
10.1111/1523-1747.ep12371797
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发表时间：
1994-03-01
影响因子：
6.5
通讯作者：
PONEC, M
中科院分区：
文献类型：
--
作者：
FARTASCH, M;PONEC, M
在气液界面培养的人角质形成细胞的表皮(包括角质层)附着在适当的底物上，其形态与体内的角质形成细胞非常相似。尽管组织学上有相似之处，但屏障功能似乎受损。本研究的目的是利用电子显微镜(包括四氧化二Ru后固定技术)和脂类成分分析来表征两种人皮肤重组体的表皮通透性屏障的发育和结构。表皮是通过在去表皮的真皮上或在含有活性成纤维细胞的牛胶原基质(相当于活皮肤)上生长的人角质形成细胞来重建的。在超微结构上，两种培养系统均表现为：(1)板层小体输送系统异常；(2)向板层脂双层转化障碍；(3)细胞间隙内表皮脂质的结构组织和分布受损。在所使用的两种系统中，培养周期的延长都没有引起角质层脂肪组织的任何显著改善。而活体皮肤仅显示稀疏的板层小体，而人角质形成细胞在常规培养液中生长的去表皮真皮上培养的板层小体数量似乎与天然皮肤相当。与活体皮肤等价物相反，脱皮真皮上的角质形成细胞培养含有更多的角膜细胞内脂滴，与脂质分析中较高的甘油三酯含量相关。通过在去表皮真皮上重建角质形成细胞培养，用与活体皮肤相同的介质，改善了脂肪组成(较低的甘油三酯含量，较高的神经酰胺含量)和结构组织，并出现了与天然皮肤相似的规则的板层脂肪双层。
The epidermis (including stratum corneum) of human keratinocytes cultured at the air-liquid interface attached to an appropriate substrate shows a morphology closely mimicking that of its in vivo counterpart. In spite of the histologic similarities, the barrier function seems to be impaired. The aim of the present study was to characterize development and structure of the epidermal permeability barrier in two human skin recombinants using electron microscopy (including ruthenium tetroxide-post fixation technique) and analysis of lipid composition. The epidermis was reconstructed by growing human keratinocytes either on de-epidermized dermis or on a bovine collagen-containing matrix with active fibroblasts (Living Skin Equivalent). Ultrastructurally both culture systems showed a) an abnormal lamellar body delivery system, b) disturbance of transformation into lamellar lipid bilayers, c) an impaired structural organization and distribution of the epidermal lipids in the intercellular spaces. In either of the systems used, prolongation of the culture period did not induce any significant improvement in the stratum corneum lipid organization. Whereas the Living Skin Equivalent showed only sparse lamellar bodies, the number of lamellar bodies in the human keratinocyte culture on de-epidermized dermis grown in regular medium seemed to be comparable to native skin. Contrary to the Living Skin Equivalent, the keratinocyte culture on de-epidermized dermis contained a higher number of intracorneocytic lipid droplets correlating with a higher triglyceride content in the lipid analyses. By reconstructing the keratinocyte culture on de-epidermized dermis with the same medium as used for the Living Skin Equivalent, both lipid composition (lower triglyceride, higher ceramide contents) and structural organization were improved, and regular lamellar lipid bilayers comparable to those of native skin appeared.