胰腺癌肝转移体外模型的构建

Establishment of a novel experimental liver metastasis model of human pancreatic cancer in vitro

目的使用旋转细胞培养系统(rotatingcellculturesystem,RCCS)模拟微重力环境,建立胰腺癌肝转移的体外模型。方法将人肝组织块和人胰腺癌细胞株BxPC3置于旋转细胞培养系统中混合培养,在额定时间点取肝组织块送病理,HE染色观察肝组织结构和胰腺癌细胞在肝组织块中生长情况。结果混合培养第1天人肝组织块周围和管腔内即有胰腺癌细胞粘附并聚集;第3天可见肝组织块中有散在癌细胞浸润;第7天可见微小转移灶形成;混合培养第10天胰腺癌肝转移灶、肝细胞形态和肝小叶结构可保持完好,并可见癌组织块形成。结论使用旋转细胞培养系统模拟微重力环境,构建胰腺癌肝转移体外模型,可以用于胰腺癌肝转移过程和机制的研究,为研究胰腺癌细胞和肝组织间的相互关系提供新的方法。

Objective To establish a novel experimental liver metastasis model of human pancreatic cancer in vitro using the rotating cell culture system (RCCS) to simulate micro-gravity culture. Methods Human normal liver tissues and human pancreatic cancer cell line BxPC-3 were co-cultured in the micro-gravity environment simulated by RCCS. The human pancreatic cancer BxPC-3 cells and human normal liver tissue explants were maintained in D-MEM/F-12 supplemented with 10% fetal calf serum and incubated at 37℃ in a 5%CO_2 incubator. The vessel was rotated at a speed of 16 rpm so that the liver tissue could be maintained in the state of suspension. The culture medium was replaced every day. The samples were taken and medium replaced under sterile conditions. The samples were stained with HE for observation of growth of pancreatic cancer cells in the liver tissue. Results The pancreatic cancer cells adhered to and grew on the liver tissue surface 1 day, massively grew in the liver tissue 7 days and formed metastatic colonies in the liver tissue 10 days after their incubation in the culture medium. Conclusions This model provides a new tool for investigating the mechanism of liver metastasis of pancreatic cancer.