Dillard Pierre, Köksal Hakan, Inderberg Else-Marit, Wälchli Sebastien
Department of Cellular Therapy, Department of Oncology, Oslo University Hospital-Radiumhospitalet.
Department of Cellular Therapy, Department of Oncology, Oslo University Hospital-Radiumhospitalet;
J Vis Exp. 2018 Dec 12(142). doi: 10.3791/58785.
Immunotherapy has become a field of growing interest in the fight against cancer otherwise untreatable. Among all immunotherapeutic methods, chimeric antigen receptor (CAR) redirected T cells obtained the most spectacular results, in particular with pediatric B-acute lymphoblastic leukemia (B-ALL). Classical validation methods of CAR T cells rely on the use of specificity and functionality assays of the CAR T cells against target cells in suspension and in xenograft models. Unfortunately, observations made in vitro are often decoupled from results obtained in vivo and a lot of effort and animals could be spared by adding another step: the use of 3D culture. The production of spheroids out of potential target cells that mimic the 3D structure of the tumor cells when they are engrafted into the animal model represents an ideal alternative. Here, we report an affordable, reliable and easy method to produce spheroids from a transduced colorectal cell line as a validation tool for adoptive cell therapy (exemplified here by CD19 CAR T cells). This method is coupled with an advanced live imaging system that can follow spheroid growth, effector cells cytotoxicity and tumor cell apoptosis.
免疫疗法已成为对抗其他无法治疗的癌症的一个越来越受关注的领域。在所有免疫治疗方法中，嵌合抗原受体（CAR）重定向T细胞取得了最显著的成果，尤其是在儿童B淋巴细胞白血病（B-ALL）方面。CAR T细胞的经典验证方法依赖于使用CAR T细胞针对悬浮靶细胞和异种移植模型中的靶细胞的特异性和功能测定。不幸的是，体外观察结果往往与体内获得的结果脱节，通过增加另一个步骤：使用3D培养，可以节省大量的精力和动物。从潜在靶细胞中产生球体，当它们植入动物模型时模仿肿瘤细胞的3D结构，这是一种理想的替代方法。在这里，我们报告了一种经济实惠、可靠且简便的方法，用于从转导的结肠直肠细胞系中产生球体，作为过继性细胞疗法的验证工具（此处以CD19 CAR T细胞为例）。该方法与先进的实时成像系统相结合，该系统可以跟踪球体生长、效应细胞的细胞毒性和肿瘤细胞凋亡。
