体外构建三维肝肿瘤模型及药物筛选

In vitro constructing a of three-dimensional hepatocarcinoma model for drug screening

背景:体外构建三维肿瘤模型替代现有二维平面肿瘤细胞模型用于药物筛选是肿瘤药筛技术发展的必然趋势。目的:体外构建三维肝肿瘤模型体系,并用于抗肿瘤药物的敏感性研究。方法:以人肝癌细胞HepG2作为模型细胞,以壳聚糖/胶原混合材料制备水凝胶支架,体外构建肝(肿瘤)细胞的三维培养体系,表征三维肝(肿瘤)细胞聚集体的形态、生长、细胞骨架分布等,并以二维平面培养的肝肿瘤细胞为对照,研究三维肝肿瘤模型对临床常用的化疗药物的敏感性。结果与结论:①肝细胞在壳聚糖/胶原水凝胶支架中培养10 d后形成三维的聚集细胞团。②肝细胞在水凝胶支架中生长速度略慢于二维平面培养,但在三维体系下肝细胞能长时间保持细胞活性。③在水凝胶支架中肝细胞三维生长后,纤维蛋白骨架发生重排,结构与在体肝组织更接近。④在水凝胶支架中的三维肝肿瘤细胞模型对化疗药物的敏感性降低。由此可见,在壳聚糖/胶原水凝胶支架中形成的三维肝(肿瘤)模型,其细胞骨架结构更接近体内肝组织,因此可用于体外药筛模型研究。

BACKGROUND：In vitro construction of three-dimensional （3D） tumor model has been growing to substitute two-dimensional （2D） tumor model for drug screening. OBJECTIVE：To develop 3D hepatocarcinoma model for the sensitive study of antitumor drugs. METHODS：Taking HepG2 as cellmodel, hydrogel scaffold was fabricated with chitosan/col agen to construct in vitro 3D hepatocarcinoma model. The 3D hepatocarcinoma aggregates were characterized regarding to the morphology, growth and cytoskeleton distribution and so on. Final y, the sensitive assay of in vitro 3D hepatocarcinoma model to the clinical antitumor drugs was studied with 2D hepatocarcinoma model as control. RESULTS AND CONCLUSION：（1） HepG2 cells in chitosan/col agen scaffold grew to form 3D cellaggregates after 10-day culture. （2） Although the growth rate of HepG2 cells in chitosan/col agen scaffold was slightly slower than that of cells in 2D culture, the HepG2 cellviability of 3D culture could be maintained longer. （3） It was found that the fibrin skeleton of HepG2 cells in chitosan/col agen scaffold rearranged and displayed structural similarity to in vivo hepatic tissue. （4） The sensitivity of in vitro 3D hepatocarcinoma model to the clinical antitumor drugs was significantly lower than that of 2D cells. In conclusion, the in vitro 3D hepatocarcinoma model developed in chitosan/col agen scaffold provided cytoskeleton structure closer to in vivo hepatic tissue, which is potential system for in vitro drug screening.