Moulin Véronique J
Centre LOEX de L'Université Laval, Génie tissulaire et régénération, LOEX, Québec, QC, Canada.
Methods Mol Biol. 2013;961:287-303. doi: 10.1007/978-1-62703-227-8_19.
Skin fibrosis is involved in several pathologies as hypertrophic scar or scleroderma. The determination of the mechanisms at the origin of these problems is however difficult due to the low number of in vivo models. To bypass this absence of animal models, studies typically use human pathological cells cultured in a monolayer way on plastic. However, cell behavior is different according to the fact that cells are on plastic or embedded in matrix. Using a tissue engineering method, we have developed new in vitro models to study these pathologies of the skin. Human pathological cells are used to reconstitute a three dimensional fibrotic tissue comprising the dermal and the epidermal parts of the skin. This method is called the self-assembly approach and is based on the cell capacity to reconstitute in vitro their own environment as in vivo. In this chapter, protocols generating reconstructed pathological skin using this approach are detailed. The methods include extraction and culture of human skin keratinocytes and fibroblasts from very small cutaneous biopsies. In addition, a description of the protocols for the production of fibrotic dermal sheets can be found to obtain a model of fibrotic dermis that can be associated or not with a fully differentiated epidermis.
皮肤纤维化涉及多种病理状况，如增生性瘢痕或硬皮病。然而，由于体内模型数量较少，确定这些问题根源的机制颇具难度。为克服动物模型缺失的问题，研究通常使用在塑料上以单层方式培养的人类病理细胞。然而，细胞行为会因细胞是在塑料上还是嵌入基质中而有所不同。我们采用组织工程方法开发了新的体外模型来研究这些皮肤病理状况。使用人类病理细胞重建包含皮肤真皮和表皮部分的三维纤维化组织。这种方法称为自组装方法，其基于细胞在体外重建自身体内环境的能力。在本章中，详细介绍了使用这种方法生成重建病理皮肤的方案。这些方法包括从非常小的皮肤活检组织中提取和培养人类皮肤角质形成细胞和成纤维细胞。此外，还可找到关于生产纤维化真皮片的方案描述，以获得可与完全分化的表皮相关联或不相关联的纤维化真皮模型。
