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使用BioStation CT进行延时成像分析:一种用于三维细胞培养的灵敏药物筛选方法。

Time-lapse imaging assay using the BioStation CT: a sensitive drug-screening method for three-dimensional cell culture.

作者信息

Sakamoto Ruriko, Rahman M Mamunur, Shimomura Manami, Itoh Manabu, Nakatsura Tetsuya

机构信息

SCIVAX Corporation, Kawasaki, Kanagawa, Japan.

SCIVAX USA, Woburn, Massachusetts, USA.

出版信息

Cancer Sci. 2015 Jun;106(6):757-765. doi: 10.1111/cas.12667. Epub 2015 May 6.

Abstract

Three-dimensional (3D) cell culture is beneficial for physiological studies of tumor cells, due to its potential to deliver a high quantity of cell culture information that is representative of the cancer microenvironment and predictive of drug responses in vivo. Currently, gel-associated or matrix-associated 3D cell culture is comprised of intricate procedures that often result in experimental complexity. Therefore, we developed an innovative anti-cancer drug sensitivity screening technique for 3D cell culture on NanoCulture Plates (NCP) by employing the imaging device BioStation CT. Here, we showed that the human breast cancer cell lines BT474 and T47D form multicellular spheroids on NCP plates and compared their sensitivity to the anti-cancer drugs trastuzumab and paclitaxel using the BioStation CT. The anticancer drugs reduced spheroid migration velocity and suppressed spheroid fusion. In addition, primary cells derived from the human breast cancer tissues B58 and B61 grown on NCP plates also exhibited similar drug sensitivity. These results were in good agreement with the conventional assay method using ATP quantification. We confirmed the antitumor effects of the drugs on cells seeded in 96-well plates using the BioStation CT imaging technique. We expect this method to be useful in research for new antitumor agents and for drug sensitivity tests in individually-tailored cancer treatments.

摘要

三维(3D)细胞培养有利于肿瘤细胞的生理学研究,因为它有可能提供大量代表癌症微环境并能预测体内药物反应的细胞培养信息。目前,凝胶相关或基质相关的3D细胞培养包含复杂的程序,常常导致实验的复杂性。因此,我们通过使用成像设备BioStation CT,开发了一种用于在纳米培养板(NCP)上进行3D细胞培养的创新型抗癌药物敏感性筛选技术。在此,我们展示了人乳腺癌细胞系BT474和T47D在NCP板上形成多细胞球体,并使用BioStation CT比较了它们对抗癌药物曲妥珠单抗和紫杉醇的敏感性。抗癌药物降低了球体迁移速度并抑制了球体融合。此外,在NCP板上生长的源自人乳腺癌组织B58和B61的原代细胞也表现出类似的药物敏感性。这些结果与使用ATP定量的传统检测方法高度一致。我们使用BioStation CT成像技术证实了药物对接种在96孔板中的细胞的抗肿瘤作用。我们期望这种方法在新型抗肿瘤药物研究以及个体化癌症治疗中的药物敏感性测试中有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0132/4471796/c946c034ca24/cas0106-0757-f1.jpg

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