Alves Arielly da Hora, Ennes do Valle Nicole Mastandrea, Yokota-Moreno Bruno Yukio, Galanciak Marta Caetano Dos Santos, Felix da Silva Keithy, Mamani Javier Bustamante, Sertie Andrea Laurato, de Oliveira Fernando Anselmo, Nucci Mariana Penteado, Gamarra Lionel Fernel
Hospital Israelita Albert Einstein, São Paulo 05529-060, Brazil.
LIM44, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo 05403-010, Brazil.
World J Stem Cells. 2025 Aug 26;17(8):108898. doi: 10.4252/wjsc.v17.i8.108898.
Glioblastoma multiforme (GBM) is the most aggressive and prevalent primary malignant brain tumor in adults, marked by poor prognosis and high invasiveness. Traditional GBM invasion assays, such as those involving mouse brain xenografts, are often time-consuming and limited in efficiency. In this context, stem cell-derived neural organoids (NOs) have emerged as advanced, three-dimensional, human-relevant platforms that mimic the cellular architecture and microenvironment of the human brain. These models provide novel opportunities to investigate glioblastoma stem cell invasion, a critical driver of tumor progression and therapeutic resistance.
To evaluate studies using stem cell-derived NOs to model glioblastoma migration/invasion, focusing on methodologies, applications and therapeutic implications.
We conducted a systematic review following PRISMA guidelines, searching PubMed and Scopus for studies published between March 2019 and March 2025 that investigated NOs in the context of glioblastoma invasion/migration. After screening 377 articles based on predefined inclusion and exclusion criteria, 10 original research articles were selected for analysis. Extracted data were categorized into four analytical domains: (1) Tumor model formation; (2) NO characteristics; (3) NO differentiation protocols; and (4) Invasion/migration assessment methodologies.
The included studies exhibit significant methodological heterogeneity GBM model development, particularly regarding model type, cell source and culture conditions. Most studies (70%) used suspension cell models, while 30% employed spheroids, with most research focusing on patient-derived glioblastoma stem cells. NOs were predominantly generated from human induced pluripotent stem cells using both guided and unguided differentiation protocols. Confocal fluorescence microscopy was the primary method used for assessing invasion, revealing invasion depths of up to 300 μm. Organoid maturity and co-culture duration influenced results, while key factors for model optimization included tumor cell density, organoid age and extracellular matrix composition. Some studies also tested therapeutic strategies such as Zika virus and microRNA modulation. Collectively, findings support the utility of NOs as effective tools for studying GBM behavior and therapeutic responses in a humanized three-dimensional context.
Human NOs represent promising platforms for modeling glioblastoma invasion in a humanized three-dimensional environment. However, a limited number of studies and methodological heterogeneity hinder reproducibility. Protocol standardization is essential to enhance the translational application of these models.
多形性胶质母细胞瘤（GBM）是成人中最具侵袭性和最常见的原发性恶性脑肿瘤，其特点是预后差和侵袭性高。传统的GBM侵袭检测方法，如涉及小鼠脑异种移植的方法，往往耗时且效率有限。在此背景下，干细胞衍生的神经类器官（NOs）已成为先进的、三维的、与人类相关的平台，可模拟人类大脑的细胞结构和微环境。这些模型为研究胶质母细胞瘤干细胞侵袭提供了新机会，而胶质母细胞瘤干细胞侵袭是肿瘤进展和治疗耐药性的关键驱动因素。
评估使用干细胞衍生的NOs模拟胶质母细胞瘤迁移/侵袭的研究，重点关注方法、应用和治疗意义。
我们按照PRISMA指南进行了系统综述，在PubMed和Scopus上搜索2019年3月至2025年3月期间发表的在胶质母细胞瘤侵袭/迁移背景下研究NOs的研究。根据预先定义的纳入和排除标准筛选了377篇文章后，选择了10篇原创研究文章进行分析。提取的数据分为四个分析领域：（1）肿瘤模型形成；（2）NO特征；（3）NO分化方案；（4）侵袭/迁移评估方法。
纳入的研究在GBM模型开发方面表现出显著的方法学异质性，特别是在模型类型、细胞来源和培养条件方面。大多数研究（70%）使用悬浮细胞模型，而30%使用球体模型，大多数研究集中在患者来源的胶质母细胞瘤干细胞。NOs主要由人诱导多能干细胞使用引导和非引导分化方案生成。共聚焦荧光显微镜是评估侵袭的主要方法，显示侵袭深度可达300μm。类器官成熟度和共培养持续时间影响结果，而模型优化的关键因素包括肿瘤细胞密度、类器官年龄和细胞外基质组成。一些研究还测试了寨卡病毒和微小RNA调节等治疗策略。总体而言，研究结果支持NOs作为在人源化三维环境中研究GBM行为和治疗反应的有效工具的实用性。
人源NOs是在人源化三维环境中模拟胶质母细胞瘤侵袭的有前景的平台。然而，研究数量有限和方法学异质性阻碍了可重复性。方案标准化对于增强这些模型的转化应用至关重要。
