Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor in adults. Despite modern, multimodal therapeutic options of surgery, chemotherapy, tumor-treating fields (TTF), and radiotherapy, the 5-year survival is below 10%. In order to develop new therapies, better preclinical models are needed that mimic the complexity of a tumor. In this work, we established a novel three-dimensional (3D) model for patient-derived GBM cell lines. To analyze the volume and growth pattern of primary GBM cells in 3D culture, a CoSeedisTMculture system was used, and radiation sensitivity in comparison to conventional 2D colony formation assay (CFA) was analyzed. Both culture systems revealed a dose-dependent reduction in survival, but the high variance in colony size and shape prevented reliable evaluation of the 2D cultures. In contrast, the size of 3D spheroids could be measured accurately. Immunostaining of spheroids grown in the 3D culture system showed an increase in the DNA double-strand-break marker γH2AX one hour after irradiation. After 24 h, a decrease in DNA damage was observed, indicating active repair mechanisms. In summary, this new translational 3D model may better reflect the tumor complexity and be useful for analyzing the growth, radiosensitivity, and DNA repair of patient-derived GBM cells.
多形性胶质母细胞瘤（GBM）是成人中最常见的恶性原发性脑肿瘤。尽管现有手术、化疗、肿瘤治疗场（TTF）及放疗等多种现代联合治疗手段，其五年生存率仍低于10%。为开发新疗法，需要能模拟肿瘤复杂性的更优临床前模型。本研究建立了一种新型三维（3D）模型用于培养患者来源的GBM细胞系。为分析原代GBM细胞在3D培养中的体积与生长模式，采用CoSeedisTM培养系统，并与传统二维克隆形成实验（CFA）对比评估其辐射敏感性。两种培养系统均显示辐射剂量依赖性的存活率下降，但二维培养因克隆大小和形态的高变异性而难以可靠评估。相比之下，3D球体的尺寸可被精确测量。对3D培养系统中生长的球体进行免疫染色显示，辐照一小时后DNA双链断裂标志物γH2AX增加，24小时后观察到DNA损伤减少，表明存在活跃的修复机制。总之，这一新型转化3D模型能更好地反映肿瘤复杂性，可用于分析患者来源GBM细胞的生长、放射敏感性与DNA修复过程。
Establishment of a 3D Model to Characterize the Radioresponse of Patient-Derived Glioblastoma Cells
肿瘤(癌症)患者之家