Yoshida Takahiro, Okuyama Hiroaki, Endo Hiroko, Inoue Masahiro
Department of Urology, The James Buchanan Brady Urological Institute, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Department of Biochemistry, Osaka International Cancer Institute, 3-1-69 Otemae, Chuo-ku, Osaka, 541-8567, Japan.
Methods Mol Biol. 2018;1655:145-153. doi: 10.1007/978-1-4939-7234-0_12.
Increasingly, it has been recognized that studying cancer samples from individual patients is important for the development of effective therapeutic strategies and in endeavors to overcome therapy resistance. Primary cultures of cancer cells acutely dissected from individual patients can provide a platform that enables the study and characterization of individual tumors. To that end, we have developed a method for preparing cancer cells in the form of multi-cellular spheroids. The cells can be derived from patient tumors (primary cells), from patient-derived xenografts, or from genetically- or chemically induced animal tumors. This method of culturing spheroids composed of cells derived from cancer tissues can be applied to various types of cancer, including urothelial cancer. The method is based on the principle of retaining cell-cell contact throughout cancer cell preparation and culturing. The first step is a partial digestion of the tumor specimen into small fragments; these fragments spontaneously form spheroidal shapes within several hours. The spheroid is referred to as a cancer tissue-originated spheroid (CTOS). The advantage of the CTOS method is that it allows one to prepare pure cancer cells at high yield. CTOSs can be stably cultured in serum-free conditions. The CTOS method can be applied to drug sensitivity assays, drug screening, and analyses of intracellular signaling. Moreover, the CTOS method provides a platform for studying the nature of cancer cell clusters.
越来越多的人认识到，研究个体患者的癌症样本对于制定有效的治疗策略以及克服治疗耐药性的努力至关重要。从个体患者身上急性解剖得到的癌细胞原代培养物可以提供一个平台，用于研究和表征个体肿瘤。为此，我们开发了一种制备多细胞球体形式癌细胞的方法。这些细胞可以来源于患者肿瘤（原代细胞）、患者来源的异种移植物，或者来源于基因或化学诱导的动物肿瘤。这种培养由癌症组织来源的细胞组成的球体的方法可以应用于各种类型的癌症，包括尿路上皮癌。该方法基于在整个癌细胞制备和培养过程中保持细胞间接触的原理。第一步是将肿瘤标本部分消化成小碎片；这些碎片在数小时内自发形成球形。这种球体被称为癌症组织起源球体（CTOS）。CTOS方法的优点是它能让人们以高产率制备纯癌细胞。CTOS可以在无血清条件下稳定培养。CTOS方法可应用于药物敏感性测定、药物筛选以及细胞内信号传导分析。此外，CTOS方法为研究癌细胞簇的性质提供了一个平台。
