三维培养的人皮肤成纤维细胞的实验研究

目的探讨三维培养的人皮肤成纤维细胞的生物学特性,为其在皮肤组织工程中的应用提供进一步的实验依据。方法培养人皮肤成纤维细胞,制备鼠尾胶原;对人皮肤成纤维细胞进行三维培养,流式细胞仪检测细胞的增殖和凋亡,RTPCR检测TGFβ1、层粘连蛋白(fibronectin,Fn)mRNA的表达,电镜观察细胞的形态学特征。结果三维培养的人皮肤成纤维细胞增殖缓慢,未见明显细胞凋亡。RTPCR检测二维、三维培养的人皮肤成纤维细胞TGFβ1、FnmRNA的表达无明显差异。电镜观察细胞染色质多,细胞器丰富。结论三维培养的人皮肤成纤维细胞增殖缓慢,但生物学活性良好。

Experimental study of human skin fibroblasts cultured in three-dimention(3D)

OBJECTIVE: To investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL). METHODS: The human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morpology. RESULTS: The flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles. CONCLUSIONS: The proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.