Zhang Mei, Boughton Philip, Rose Barbara, Lee C Soon, Hong Angela M
Department of Radiation Oncology, Royal Prince Alfred Hospital, Sydney, NSW 2050, Australia ; Sydney Medical School, The University of Sydney, Sydney, NSW 2006, Australia.
Int J Biomater. 2013;2013:396056. doi: 10.1155/2013/396056. Epub 2013 Sep 11.
Background. Human cancer is a three-dimensional (3D) structure consisting of neighboring cells, extracellular matrix, and blood vessels. It is therefore critical to mimic the cancer cells and their surrounding environment during in vitro study. Our aim was to establish a 3D cancer model using a synthetic composite scaffold. Methods. High-density low-volume seeding was used to promote attachment of a non-small-cell lung cancer cell line (NCI-H460) to scaffolds. Growth patterns in 3D culture were compared with those of monolayers. Immunohistochemistry was conducted to compare the expression of Ki67, CD44, and carbonic anhydrase IX. Results. NCI-H460 readily attached to the scaffold without surface pretreatment at a rate of 35% from a load of 1.5 × 10(6) cells. Most cells grew vertically to form clumps along the surface of the scaffold, and cell morphology resembled tissue origin; 2D cultures exhibited characteristics of adherent epithelial cancer cell lines. Expression patterns of Ki67, CD44, and CA IX varied markedly between 3D and monolayer cultures. Conclusions. The behavior of cancer cells in our 3D model is similar to tumor growth in vivo. This model will provide the basis for future study using 3D cancer culture.
背景。人类癌症是一种三维（3D）结构，由相邻细胞、细胞外基质和血管组成。因此，在体外研究期间模拟癌细胞及其周围环境至关重要。我们的目的是使用合成复合支架建立一种3D癌症模型。方法。采用高密度低体积接种法促进非小细胞肺癌细胞系（NCI-H460）附着于支架。将3D培养中的生长模式与单层培养的生长模式进行比较。进行免疫组织化学以比较Ki67、CD44和碳酸酐酶IX的表达。结果。NCI-H460在未经表面预处理的情况下易于附着于支架，接种1.5×10⁶个细胞时附着率为35%。大多数细胞垂直生长，沿支架表面形成团块，细胞形态类似于组织来源；二维培养表现出贴壁上皮癌细胞系的特征。Ki67、CD44和CA IX的表达模式在3D培养和单层培养之间有明显差异。结论。我们的3D模型中癌细胞的行为类似于体内肿瘤生长。该模型将为未来使用3D癌症培养的研究提供基础。
