脱细胞猪角膜基质体外支持皮肤细胞生长的实验研究

Experimental study on acellular porcine corneal stroma supporting growth of skin cells in vitro

目的探讨脱细胞猪角膜基质体外是否支持皮肤细胞的生长。方法制备脱细胞猪角膜基质(前期实验已完成)。体外培养人皮肤表皮细胞和成纤维细胞,取第3代人皮肤成纤维细胞接种在脱细胞猪角膜基质的中间基质面,培养3d后,将材料翻转,取第3代人皮肤表皮细胞接种在材料的上皮面上,再培养10d后,取细胞-支架复合体制作石蜡切片,HE染色后光镜下观察。结果组织学观察显示皮肤的表皮细胞和成纤维细胞体外均可在脱细胞猪角膜基质上黏附生长。接种10d后,皮肤表皮细胞在材料表面形成复层结构,可见角化的细胞。接种13d可见成纤维细胞存活并在支架层间生长。结论脱细胞猪角膜基质有良好的细胞相容性,在体外能支持皮肤细胞的生长和增生。

Objective To determine acellular porcine corneal stroma （APCS） supporting the growth of skin cells in vitro or not. Methods APCS had been prepared at pre-experiment. The epidermic cells and fibroblasts of human skin were cultured in vitro. The 3rd passaged fibroblasts of human skin were seeded on the middle stromal side of APCS. After cultured for 3 days, the cell-APCS constructs were turned over and the 3rd passaged epidermic cells of human skin were seeded onto the epithelium side of APCS. After cultured for another 10 days, the cell-scaffold constructs were taken paraffin section, and observed by light microscopy after HE staining. Results The histological examination showed the epidermic cells and the fibroblasts of human skin could grow adhesively on APCS in vitro. The epidermic cells of the skin grew in multi-layers at 10 days after inoculation. Some cells presented keratinization. The fibroblasts were alive and invaded in the interlaminar of the scaffold at 13 days after inoculation. Conclusion APCS has a good cell compatibility and can support the growth and proliferation of skin cells in vitro.