患者肿瘤组织来源的胶质母细胞瘤类器官的制备方法

目的：探讨胶质母细胞瘤（GBM）患者肿瘤组织来源的GBM类器官（GBO）模型的制备方法。方法：选取2021年广东三九脑科医院新诊断经病理确诊的8例GBM患者的新鲜肿瘤组织标本，将其剪成0.5～1 mm大小的组织碎片并用特制的培养基进行培养，待其成球且直径达到1 mm时剪小传代，同时选取培养2周以上的GBO进行石蜡包埋、切片，后进行H-E染色和免疫组化染色检测，并与亲本GBM组织进行组织学与细胞学的比较。结果：成功培养2例可传代冻存的GBO，并建立GBO生物库。H-E染色结果显示，GBO保留了与亲本GBM组织相似的组织结构和细胞形态；免疫组化实验结果显示，GBO与GBM组织中GFAP、OLIG2、Ki67和ATRX分子的表达情况一致。结论：将患者来源的GBM组织在体外剪小并用特制培养基培养，可构建与GBM患者肿瘤组织在组织和细胞层面一致的GBO。

A preparation method for the organoid model of patient-derived glioblastoma

Objective: To explore a preparation method for glioblastoma organoid (GBO) derived from patients with glioblastoma (GBM). Methods: The fresh tumor tissue samples of eight GBM patients newly diagnosed and pathologically confirmed in Guangdong 999 Brain Hospital in 2021 were selected, cut into tissue fragments of 0.5-1 mm in size and cultured in a special medium. When the tissue fragments grew into a spherical shape and the diameter reached 1 mm, they were cut into small pieces and passaged. GBOs cultured for more than 2 weeks were selected for paraffin-embedding, sectioned, and then stained with H-E and immunohistochemical staining. Histological and cytological comparisons were performed with parental GBM tissues. Results: Two cases of GBO that could be passaged and cryopreserved were successfully prepared and a GBO biobank was established. The results of H-E staining showed that GBOs retained tissue structure and cell morphology similar to those of the parental GBM tissues. The results of immunohistochemical analysis showed that the expressions of GFAP, OLIG2, Ki67 and ATRX in GBO and parental GBM tissues were basically consistent. Conclusion: GBO can be generated in vitro by trimming patient-derived GBM tissue and culturing it in a suitable medium. The GBO prepared by this method is consistent with the parental GBM tissue at the histological and cytological levels.