应用改良小室法构建裸鼠皮肤及毛囊

Application of modified chamber method on reconstituting skin and hair follicle of nude mouse

目的应用改良小室法在祼鼠体内构建出有正常组织结构的皮肤和毛发。方法应用16.5 d小鼠胚胎的表皮细胞、成纤维细胞和Ⅰ型鼠尾胶原蛋白构建毛囊重建模型。首先在裸鼠背部做直径1.0 cm的圆形创面,将小室固定于创面上;然后,在小室内加入成纤维细胞和Ⅰ型鼠尾胶原蛋白混合悬液形成真皮层,等待胶原凝固后加入表皮细胞形成表皮层。对照组采用传统小室法,16.5 d小鼠胚胎的表皮细胞和成纤维细胞混匀后加入裸鼠体内构建好的小室中。结果实验组和对照组在第10天拆除小室后创面皮肤均得到修复,两组无明显差异。拆除小室1周后,实验组在创面上形成均匀分布的毛发,对照组毛发呈现不均匀分布。拆除小室1个月后可观察到实验组和对照组创面均能够形成大量毛发。通过HE染色及免疫荧光检测发现改良小室法所构建的皮肤拥有正常的组织结构及完整的毛囊结构。结论改良小室法能够构建出有正常组织结构的皮肤组织,是一个高效稳定的毛囊重建模型。

Objective Application of modifid chamber method contained normal tissue structure of mouse skin and completed hair follicle in nude mice. Methods Epidermal cells and fibroblasts from 16.5 days embryonic mice,and collagen type I rat tail were applied to construct hair follicle reconstitution model. First, the skin on the back of nude mouse was removed to create a hole of 10 mm in diameter, and a chamber was fixed to the hole. Then a dermis layer built up by fibroblasts and collagen type I from rat tail was implanted into the chamber. Epidermal cells were added once the collagen solidified. Results Both the test group and control group had skin repaired, and there was no significant difference at10 days after surgery. One week after chamber removal, the wound of test group generated evenly distributed hair while the regenerated hair control group was non-evenly distributed. After removed the chamber one month, both test group and control group could be observed with large amount of hair generation on the wound.Application of modified chamber method constructed normal tissue structure of the skin and hair, through HE staining and immunofluorescence detection.This method was able to generate new skin, which contained normal tissue structure of mouse skin and completed hair follicle function. Conclusion Modified chamber method is an efficient and stable follicle reconstitution model.