发展3 d球体分析高温超导药物发现方法

抑制肿瘤细胞增殖的药物发现研究翻译不好从传统的二维(2 d)在体内体外实验研究。执行大多数抗癌药物化合物研究组织培养处理二维分析格式研究扩散的目的,生存能力和细胞凋亡。越来越多的科学证据表明癌细胞增长的三维(3 d)球状体体内研究的预测结果比2 d细胞培养格式。通常,使用各种方法创建球状体,如U-bottom盘子,悬滴或半固体媒体和分析领域的增长进行了镜下,费时、费力、低吞吐量和缺乏再现性。文献报道的使用一个自动化的成像仪监测三维球状体的大小和增长在96 -孔板,在球体形成4天从一组细胞的数量返回一个球体大小一致。在这项研究中,我们强调使用384 -低依恋U-bottom板结合Celigo®成像血细胞计数器图像和分析三维球状体的形成,允许增加吞吐量,每个板块的复制和参数数量相比96 -孔板。U87MG tumorspheres创建在384 -孔板,随后分析了成像。tumorspheres被监控的21天成像仪报道从brightfield图像定量测量,包括大小和动力学领域的扩散。此外,附着创建2 d盘子比较。Z”因素的鲁棒性和IC50值报告(Z的0.6倍为3 d和2 d分析格式,IC50 0.17 0.25嗯,嗯,分别),说明可再生的3 d在384孔板可以形成球状体。 Fluorescent studies were carried out with the imager for viability using intensity analysis. For compound screening methods, the assay was evaluated in comparison using various drug compounds that have shown anti-proliferative effects. Similarly reported with 17AAG data, comparison of IC50 data reported higher values in the 3D assay than in the 2D assay, (doxorubicin: 3D, 0.166uM and 2D, 0.049uM; TMZ: 3D, 15uM and 2D, 13uM), except in the paclitaxel response, where 0.012uM was reported for both assays. Together, these data demonstrate that the tumorsphere formation assay can be developed, validated and used for high-throughput anti-cancer compound screening in 384-well U-Boxall Fbottom low attachment plates using the Celígo Imaging Cytometer.