Celoptics. Inc, Rockville, MD, USA.
Department of Neurological Surgery, Mayo Clinic, Jacksonville, FL, USA.
J Neurooncol. 2023 Aug;164(1):43-54. doi: 10.1007/s11060-023-04349-9. Epub 2023 Jul 25.
Glioblastoma (GBM) is an aggressive primary brain cancer. Lack of effective therapy is related to its highly invasive nature. GBM invasion has been studied with reductionist systems that do not fully recapitulate the cytoarchitecture of the brain. We describe a human-derived brain organotypic model to study the migratory properties of GBM IDH-wild type ex vivo.
Non-tumor brain samples were obtained from patients undergoing surgery (n = 7). Organotypic brain slices were prepared, and green fluorescent protein (GFP)-labeled primary human GBM IDH-wild type cells (GBM276, GBM612, GBM965) were placed on the organotypic slice. Migration was evaluated via microscopy and immunohistochemistry.
After placement, cells migrated towards blood vessels; initially migrating with limited directionality, sending processes in different directions, and increasing their speed upon contact with the vessel. Once merged, migration speed decreased and continued to decrease with time (p < 0.001). After perivascular localization, migration is limited along the blood vessels in both directions. The percentage of cells that contact blood vessels and then continue to migrate along the vessel was 92.5% (- 3.9/ + 2.9)% while the percentage of cells that migrate along the blood vessel and leave was 7.5% (- 2.9/ + 3.9) (95% CI, Clopper-Pearson (exact); n = 256 cells from six organotypic cultures); these percentages are significantly different from the random (50%) null hypothesis (z = 13.6; p < 10). Further, cells increase their speed in response to a decrease in oxygen tension from atmospheric normoxia (20% O) to anoxia (1% O) (p = 0.033).
Human organotypic models can accurately study cell migration ex vivo. GBM IDH-wild type cells migrate toward the perivascular space in blood vessels and their migratory parameters change once they contact vascular structures and under hypoxic conditions. This model allows the evaluation of GBM invasion, considering the human brain microenvironment when cells are removed from their native niche after surgery.
胶质母细胞瘤（GBM）是一种侵袭性原发性脑癌。缺乏有效的治疗方法与其高度侵袭性有关。GBM 的侵袭性已通过简化系统进行了研究，但这些系统不能完全再现大脑的细胞结构。我们描述了一种人源性脑器官型模型，用于研究 IDH 野生型 ex vivo 胶质母细胞瘤的迁移特性。
从接受手术的患者（n=7）中获得非肿瘤脑组织样本。制备器官型脑切片，并将绿色荧光蛋白（GFP）标记的原发性人 GBM IDH 野生型细胞（GBM276、GBM612、GBM965）放置在器官型切片上。通过显微镜和免疫组织化学评估迁移。
放置后，细胞向血管迁移；最初迁移方向有限，向不同方向发出突起，并在与血管接触时增加速度。一旦融合，迁移速度就会降低，并且随着时间的推移继续降低（p<0.001）。在血管周围定位后，向两个方向沿血管的迁移受到限制。接触血管然后继续沿血管迁移的细胞百分比为 92.5%（-3.9/+2.9）%，而沿血管迁移并离开的细胞百分比为 7.5%（-2.9/+3.9）%（95%置信区间，Clop-per-Pearson（精确）；n=256 个细胞来自六个器官型培养物）；这些百分比与随机（50%）零假设显著不同（z=13.6；p<10）。此外，细胞在从大气正常氧（20%O）到缺氧（1%O）的氧张力降低时会增加速度（p=0.033）。
人器官型模型可以准确地在体外研究细胞迁移。GBM IDH 野生型细胞向血管的血管周围空间迁移，并且一旦与血管结构接触并且在缺氧条件下，它们的迁移参数就会发生变化。该模型允许在考虑到手术切除后细胞从其天然生态位中取出时的人脑微环境的情况下评估 GBM 的侵袭性。
