A modified ex vivo skin organ culture system for functional studies
A modified ex vivo skin organ culture system for functional studies
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DOI：
10.1007/s004030100219
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发表时间：
2001-04-01
影响因子：
3
通讯作者：
Prens, EP
中科院分区：
文献类型：
--
作者：
Companjen, AR;van der Wel, LI;Prens, EP
为了在接近体内情况的条件下研究正常和患病皮肤中细胞的免疫功能，必须保持组织的结构完整性。为了实现这一点，必须离体培养新鲜分离的皮肤，或者可以使用体外构建的完整皮肤等同物。已经描述了不同的皮肤器官培养系统。基本上有两种系统占主导地位：浸没或空气暴露的皮肤器官培养。前者已被用于测量细胞因子分泌的皮肤细胞在培养基中，和后者研究细胞膜蛋白的表达在原位和表皮朗格汉斯细胞的动力学。在这里，我们提出了一种改进的离体皮肤器官培养系统，该系统通过维持正常的皮肤结构而不自发诱导再生成熟标志物来接近体内情况。该方法允许原位测量细胞膜蛋白的表达，并在同一实验中定量培养基中皮肤细胞的细胞因子分泌。在该系统中，一般和特异性刺激（LPS和IL-1 β）均上调培养基中皮肤来源的细胞因子IL-8和IL-6的表达，以及24小时培养形成的组织中细胞上的不同标志物（ICAM-1、CD 40和CD 86）。细胞因子和细胞标志物表达的升高可被地塞米松和特异性作用于IL-1 β介导的应答的IL-1 ra阻断。这里介绍的系统既快速又简单，可以用作研究皮肤细胞在自然微环境中行为的模型。
To investigate the immunological function of cells in normal and diseased skin under conditions approximating the in vivo situation, it is necessary to maintain the structural integrity of the tissue. To achieve this, freshly isolated skin has to be cultured ex vivo, or an in vitro-constructed complete skin equivalent may be used. Different skin organ culture systems have been described. Basically two systems prevail: submerged or air-exposed skin organ cultures. The former model has been used for measuring cytokine secretion by skin cells in the medium, and the latter to study the expression of cell membrane proteins in situ and the kinetics of epidermal Langerhans cells. Here we present a modified ex vivo skin organ culture system which approaches the in vivo situation by maintaining the normal skin architecture without spontaneous induction of the regenerative maturation markers. This method allowed the expression of cell membrane proteins in situ to be measured, and the cytokine secretion by skin cells in the culture medium to be quantitated in the same experiment. In this system, both general and specific stimuli (LPS and IL-1 beta) upregulated the expression of skin-derived cytokines IL-8 and IL-6 in the medium and different markers (ICAM-1, CD40 and CD86) on cells in the tissue in a 24-hour culture-formed. Elevation of both cytokine and cell marker expression could be blocked by dexamethasone and by IL-1ra which acts specifically on IL-1 beta -mediated responses. The system presented here is both quick and simple and can be used as a model to studs the behaviour of skin cells in their natural microenvironment.