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Updated: Nov 22, 2025
A 3D Spheroid Model for Glioblastoma
Published on: April 9, 2020
Brain-Tumor-Regenerating 3D Scaffold-Based Primary Xenograft Models for Glioma Stem Cell Targeted Drug Screening
Kottarapat Jeena1, Cheripelil Abraham Manju2, Koythatta Meethalveedu Sajesh3
1Amrita Centre for Nanosciences and Molecular Medicine, ‡Central Lab Animal Facility, and §Department of Pathology, Amrita Vishwa Vidyapeetham, Amrita Institute of Medical Sciences, Ponekkara, Kochi 682 041, India.
Abstract:
Glioma stem cells (GSC) present a critical therapeutic challenge for glioblastoma multiforme (GBM). Drug screening against GSC demands development of novel in vitro and in vivo platforms that can mimic brain microenvironment and support GSC maintenance and tumorigenesis. Here, we report, a 3-dimensionel (3D) biomimetic macro-porous scaffold developed by incorporating hyaluronic acid, porcine brain extra cellular matrix (ECM) and growth factors that facilitates regeneration of GBM from primary GSCs, ex vivo and in vivo. After characterizing with human and rat GBM cell lines and neurospheres, human GSCs expressing Notch1, Sox-2, Nestin, and CD133 biomarkers were isolated from GBM patients, cultured in the 3D scaffold, and implanted subcutaneously in nude mice to develop patient derived xenograft (PDX) models. Aggressive growth pattern of PDX with formation of intratumoral vascularization was monitored by magnetic resonance imaging (MRI). Histopathological and phenotypial features of the original tumors were retained in the PDX models. We used this regenerated GBM platform to screen novel siRNA nanotherapeutics targeting Notch, Sox-2, FAK signaling for its ability to inhibit the tumorigenic potential of GSCs. Current clinical drug, Temozolomide and an anticancer phytochemical, nanocurcumin, were used as controls. The siRNA nanoparticles showed excellent efficacy in inhibiting tumorigenesis by GSCs in vivo. Our study suggests that the brain-ECM mimicking scaffold can regenerate primary gliomas from GSCs in vitro and in vivo, and the same can be used as an effective platform for screening drugs against glioma stem cells.
Insights
A novel 3D scaffold mimics the brain microenvironment to regenerate glioblastoma and test therapies. This platform effectively screened siRNA nanotherapeutics, showing promise for glioma stem cell drug discovery.
Area of Science:
- Biomaterials Science
- Neuro-oncology
- Regenerative Medicine
Background:
- Glioblastoma stem cells (GSCs) are key therapeutic targets due to their role in glioblastoma multiforme (GBM) recurrence.
- Effective drug screening requires platforms that replicate the brain's microenvironment and support GSC maintenance and tumorigenesis.
- Current in vitro and in vivo models often fail to fully capture GSC behavior and GBM complexity.
Purpose of the Study:
- To develop a 3D biomimetic scaffold that supports GSC maintenance and tumorigenesis.
- To establish a platform for ex vivo and in vivo regeneration of GBM from patient-derived GSCs.
- To utilize this platform for screening novel nanotherapeutics against GSCs.
Main Methods:
- Fabrication of a 3D macro-porous scaffold using hyaluronic acid, porcine brain extracellular matrix (ECM), and growth factors.
- Isolation and culture of human GSCs expressing key biomarkers (Notch1, Sox-2, Nestin, CD133).
- Generation of patient-derived xenograft (PDX) models by implanting GSCs in a 3D scaffold into nude mice, monitored by MRI.
- Screening of siRNA nanotherapeutics targeting Notch, Sox-2, and FAK signaling pathways.
Main Results:
- The 3D scaffold successfully supported the regeneration of GBM from GSCs, both ex vivo and in vivo.
- PDX models exhibited aggressive growth and vascularization, retaining original tumor characteristics.
- siRNA nanotherapeutics demonstrated significant efficacy in inhibiting GSC tumorigenesis within the scaffold platform.
- The scaffold platform proved effective for screening anti-GSC therapies.
Conclusions:
- A novel brain-ECM mimicking 3D scaffold can regenerate primary gliomas from GSCs.
- This platform is effective for in vitro and in vivo drug screening against glioma stem cells.
- The developed platform holds potential for advancing therapeutic strategies for glioblastoma.
