Purpose <p>Glioblastoma multiforme is an aggressive, potentially fatal form of brain cancer characterised by an extremely poor prognosis. Glioblastoma spheroids can recapitulate the major three-dimensional tumour characteristics, encompassing cell-cell interactions, extracellular matrix deposition, and diffusion gradients for in vitro assessment. Furthermore, insufficient characterisation, inconsistent reproducibility, and inadequately standardised protocols restrict the use of spheroids as substitutes for in vivo models.</p> Methods <p>Here, we propose the fabrication of a polycaprolactone scaffold using a 3D printing system that can induce spheroid formation when seeded with glioblastoma cells. In this current study, U87-MG cells were seeded on 3D printed polycaprolactone scaffolds. The printed scaffolds containing spheroids were assessed for biocompatibility using Live/Dead staining, ATP-based viability assay, and analysis of GFAP expression. To demonstrate the practicability of the printed scaffold as a model for drug screening, U87-MG spheroids were exposed to temozolomide, and the cell viability was quantified.</p> Results <p>U87-MG spheroid formation was observed within 24&#xa0;h, indicating the suitability of 3D printed PCL scaffolds for spheroid generation. Live/Dead staining and luminescent ATP-based viability assays confirmed the biocompatibility of the scaffold and its capacity to support cellular proliferation. Analysis of GFAP expression confirmed the stemness of the U87-MG spheroids formed in the 3D printed PCL scaffolds. The spheroids were susceptible to temozolomide, with pronounced cell death and loss of architecture, as observed on the 7th day. The comparable drug response profiles observed in all three scaffold groups further support the use of 3D printed PCL scaffolds as a viable platform for drug screening.</p> Conclusion <p>3D printed PCL scaffold supported the formation of U87-MG spheroids without the need for external cues and presents a robust platform for drug screening applications.</p>

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3D-Printed Scaffold-Based Glioblastoma Spheroid In Vitro Model for Drug Screening Application

  • Iyer Aakash Sambamoorthy,
  • Bhuvaneshwari Arumugam,
  • Ceera Manikandan,
  • Amit Kumar Jaiswal

摘要

Purpose

Glioblastoma multiforme is an aggressive, potentially fatal form of brain cancer characterised by an extremely poor prognosis. Glioblastoma spheroids can recapitulate the major three-dimensional tumour characteristics, encompassing cell-cell interactions, extracellular matrix deposition, and diffusion gradients for in vitro assessment. Furthermore, insufficient characterisation, inconsistent reproducibility, and inadequately standardised protocols restrict the use of spheroids as substitutes for in vivo models.

Methods

Here, we propose the fabrication of a polycaprolactone scaffold using a 3D printing system that can induce spheroid formation when seeded with glioblastoma cells. In this current study, U87-MG cells were seeded on 3D printed polycaprolactone scaffolds. The printed scaffolds containing spheroids were assessed for biocompatibility using Live/Dead staining, ATP-based viability assay, and analysis of GFAP expression. To demonstrate the practicability of the printed scaffold as a model for drug screening, U87-MG spheroids were exposed to temozolomide, and the cell viability was quantified.

Results

U87-MG spheroid formation was observed within 24 h, indicating the suitability of 3D printed PCL scaffolds for spheroid generation. Live/Dead staining and luminescent ATP-based viability assays confirmed the biocompatibility of the scaffold and its capacity to support cellular proliferation. Analysis of GFAP expression confirmed the stemness of the U87-MG spheroids formed in the 3D printed PCL scaffolds. The spheroids were susceptible to temozolomide, with pronounced cell death and loss of architecture, as observed on the 7th day. The comparable drug response profiles observed in all three scaffold groups further support the use of 3D printed PCL scaffolds as a viable platform for drug screening.

Conclusion

3D printed PCL scaffold supported the formation of U87-MG spheroids without the need for external cues and presents a robust platform for drug screening applications.