Comparative analysis of tumor spheroid generation techniques for differential in vitro drug toxicity.
Comparative analysis of tumor spheroid generation techniques for differential in vitro drug toxicity.
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DOI：
10.18632/oncotarget.7659
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发表时间：
2016-03-29
期刊：
影响因子：
--
通讯作者：
Mehta G
中科院分区：
文献类型：
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作者：
Raghavan S;Mehta P;Horst EN;Ward MR;Rowley KR;Mehta G
多细胞肿瘤球体是进行临床前化疗敏感性测定的有效体外模型。我们比较了不同的方法来产生肿瘤球体的球体形态，细胞排列和化疗敏感性。我们使用两种癌细胞系（MCF 7和OVCAR 8）使用i）悬滴阵列板; ii）超低附着板上的液体覆盖物; iii）具有旋转混合的超低附着板（章动板）上的液体覆盖物来产生球状体。球状体形态计量学分析表明，在章动和悬滴阵列板上产生的球状体中细胞致密化增加。胶原染色还表明，与传统的液体覆盖相比，章动器和悬滴阵列上的肿瘤球体中的压实和重塑更高。因此，与超低附着板上的球状体（10-20%活力）相比，章动或悬滴板上产生的球状体对顺铂治疗的化学抗性增加（20-60%活力）。最后，我们使用了一个数学模型来证明实验生成的球体内的氧和顺铂扩散的最小变化。我们的研究结果表明，在体外方法的肿瘤球体产生不同的细胞排列和化疗敏感性。
Multicellular tumor spheroids are powerful in vitro models to perform preclinical chemosensitivity assays. We compare different methodologies to generate tumor spheroids in terms of resultant spheroid morphology, cellular arrangement and chemosensitivity. We used two cancer cell lines (MCF7 and OVCAR8) to generate spheroids using i) hanging drop array plates; ii) liquid overlay on ultra-low attachment plates; iii) liquid overlay on ultra-low attachment plates with rotating mixing (nutator plates). Analysis of spheroid morphometry indicated that cellular compaction was increased in spheroids generated on nutator and hanging drop array plates. Collagen staining also indicated higher compaction and remodeling in tumor spheroids on nutator and hanging drop arrays compared to conventional liquid overlay. Consequently, spheroids generated on nutator or hanging drop plates had increased chemoresistance to cisplatin treatment (20-60% viability) compared to spheroids on ultra low attachment plates (10-20% viability). Lastly, we used a mathematical model to demonstrate minimal changes in oxygen and cisplatin diffusion within experimentally generated spheroids. Our results demonstrate that in vitro methods of tumor spheroid generation result in varied cellular arrangement and chemosensitivity.