胆管癌细胞体外三维培养模型建立与二维培养体系中生长代谢的比较

Comparison of growth and metabolism of cholangiocarcinoma cells in two-dimensional and three-dimensional culture systems

背景:二维细胞培养是目前体外研究恶性肿瘤和检测抗肿瘤药物的常用方法,但存在局限性。目的:建立胆管癌细胞体外三维培养模型,比较在二维和三维培养体系中胆管癌细胞生长及代谢的差异。方法:将胆管癌细胞株QBC939培养于二维培养体系及Cytodex-3或Cytopore-2的三维培养体系中,观察细胞形态和数量,测定培养上清液中葡萄糖、乳酸浓度,计算细胞生长代谢动力学参数。结果与结论:与二维和Cytodex-3三维培养体系相比,由Cytopore-2构成的三维培养体系中细胞在微载体上贴壁率较高,分布均匀,所达到的细胞密度最高,且细胞的葡萄糖比消耗速率和乳酸比生成速率明显低于二维和Cytodex-3三维培养系统(P<0.05)。结果证实,采用Cytodex-3或Cytopore-2微载体培养体系均可建立胆管癌细胞的三维培养模型,且Cytopore-2三维培养体系优于二维和Cytodex-3三维培养体系。

BACKGROUND：Two-dimensional（2D） culture model is a common tool for tumor research and drug screening in vitro;however,there are some limitations.OBJECTIVE：To establish a three-dimensional（3D） culture model of cholangiocarcinoma cells and to compare the difference in cell growth and metabolism between 2D and 3D culture systems.METHODS：Cholangiocarcinoma cell line QBC939 was cultured in 2D and Cytodex-3 or Cytopore-2 microcarrier-based 3D culture systems.Cell morphology was observed under inverted light microscopy.Total cell number,glucose and lactate concentrations were determined,and the kinetic constants of cell growth and metabolism were calculated.RESULTS AND CONCLUSION：In comparison with 2D and Cytodex-3-based 3D culture systems,Cytopore-2-based 3D culture system achieved higher cell adhesion rate,higher cell density,and lower specific glucose consumption and specific lactate production rates（P〈0.05） .In addition,cells were uniformly distributed in Cytopore-2 microcarriers.Therefore,Cytodex-3 or Cytopore-2 microcarriers can be used to establish cholangiocarcinoma 3D culture model,and Cytopore-2 3D culture system is better than Cytodex-3 system.