Stem cell-derived neural organoids as platforms to investigate glioblastoma invasion and migration: A systematic review

Abstract

BACKGROUND

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.

AIM

To evaluate studies using stem cell-derived NOs to model glioblastoma migration/invasion, focusing on methodologies, applications and therapeutic implications.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

Keywords: Glioblastoma; Stem cell; Organoid; Spheroid; Invasion; Migration

Core Tip: This systematic review highlights human stem cell-derived neural organoids as promising three-dimensional models for investigating glioblastoma stem cell invasion/migration. Despite considerable methodological heterogeneity, the studies demonstrate the potential of these models to replicate key aspects of the tumor microenvironment, assess therapeutic responses, and support personalized medicine approaches. However, the lack of standardized protocols and evaluation methods poses a challenge to reproducibility and broader translational use. Standardizing methodologies will be key to advancing these models and establishing their value in translational glioblastoma research and the development of targeted therapies.