摘要
目的体外观察成体肝干细胞与相互隔离的肝癌细胞在共培养体系中的生物学行为。方法利用重组腺病毒介导,建立工程化稳定表达增强型绿色荧光蛋白(EGFP)的大鼠成体肝干细胞WB-EGFP 胚胎成纤维细胞REF EGFP。在相互隔离的体系下,共同培养肝癌细胞CBRH 7919、肝干细胞WB-EGFP 或成纤维细胞REF EGFP,连续动态观察肝干细胞在体外共培养体系中的迁移情况。结果隔离区细胞爬片上培养的WB-EGFP细胞缓慢向肝癌细胞CBRH 7919培养区移动,不仅发生在中央空白区接种的WB- EGFP细胞,而且最边缘空白区接种的WB-EGFP细胞也有类似的定向迁移运动;而成纤维细胞无趋向肿瘤细胞迁移的现象,72h后仍大部分停留在盖玻片上。结论成体肝干细胞在体外具有追踪趋向肝癌细胞的生物学特性。
Objective To explore the biological behavior of adult liver stem cells in a co-cultured system of them with hepatocelluar carcinoma （HCC） cells without direct contact between the two kinds of cells. Methods WB-F344, a kind of rat adult liver stem cell, and rat embryonic fibroblasts （REF） from a primary culture were engineered to express enhanced green fluorescent protein （EGFP） by recombinant adenoviral-mediated methods. Two kinds of cells marked with EGFP were established, namely WB-EGFP and REF-EGFP. After the HCC cells grew to 40%-60% confluence in the culture dish with a 10-mm cell-free area, a similar number of WB-EGFP and REF-EGFP were placed in the blank areas respectively. Then, we constantly studied the movement behavior of WB-EGFP in the co-culture system with HCC cells. Results The results showed that WB-EGFP cells migrated to the area of HCC ceils slowly. Their appearance was found not only when WB-EGFP cells were seeded into the cell-free area at the center of the dish, but also when seeded into the blank area at the extreme edge of the plate. This trait was not observed in the co-cultured system of REF-EGFP and HCC cells, and most REF-EGFP were still localized in their initial area after 72 hours of incubation. Conclusion The results mean that adult liver stem cells have a biological behavior of selective tropism toward HCC cells in vitro, and suggest a possibility of using migratory liver stem ceils as a delivery vehicle for gene therapy for HCC.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2005年第9期644-647,共4页
Chinese Journal of Hepatology
