A human co‐culture cell model incorporating microglia supports glioblastoma growth and migration, and confers resistance to cytotoxics

Abstract

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. AbstractDespite the importance of the tumor microenvironment in regulating tumor progression, few in vitro models have been developed to understand the effects of nonneoplastic cells and extracellular matrix (ECM) on drug resistance in glioblastoma (GBM) cells. Using CellTrace-labeled human GBM and microglial (MG) cells, we established a 2D co-culture including various ratios of the two cell types. Viability, proliferation, migration, and drug response assays were carried out to assess the role of MG. A 3D model was then established using a hyaluronic acid-gelatin hydrogel to culture a mixture of GBM and MG and evaluate drug resistance. A contact coculture of fluorescently labeled GBM and MG demonstrated that MG cells modestly promoted tumor cell proliferation (17%-30% increase) and greater migration of GBM cells (>1.5-fold increase). Notably, the presence of MG elicited drug resistance even when in a low ratio (10%-20%) relative to co-cultured tumor cells. The protective effect of MG on GBM was greater in the 3D model (>100% survival of GBM when challenged with cytotoxics). This new 3D human model demonstrated the influence of non-neoplastic cells and matrix on chemoresistance of GBM cells to three agents with different mechanisms of action suggesting that such sophisticated in vitro approaches may facilitate improved preclinical testing. K E Y W O R D S3D co-culture, drug resistance, human serum, hyaluronic acid hydrogel | 1711 LEITE ET aL.

Considering the Experimental Use of Temozolomide in Glioblastoma Research

“…Primary microglia cultures as well as tumour-associated or 'normal' endothelial cells seem to be resistant towards TMZ treatment, too [163,164]. Microglial cells have been associated with stimulating proliferation and invasion of GB cells, as well as mediating resistance towards cytotoxic drugs, such as vincristine and TMZ [165]. Co-culturing of GB cells with endothelial cells as well as stromal cells, which are important constituents of the perivascular niche of the tumour [166], led to upregulation of genes involved in angiogenesis, resulted in enhanced PI3K/Akt and Ras/MAPK signalling in GB cells and increased their resistance towards TMZ treatment [167].…”

Identification of a Metabolism-Related Risk Signature Associated With Clinical Prognosis in Glioblastoma Using Integrated Bioinformatic Analysis

“…Interestingly, our results suggest that some conventional antipsychotics among drugs mentioned above, including amitriptyline, fluoxetine, and imipramine, may also play a surprising role in the treatment of GBM, which is consistent with previous studies and offers new hope for patients with GBM (12,75,76,80,83,84,104). In addition, Leite et al (105) demonstrated that clomipramine (tricyclic antidepressant drugs such as imipramine) has an impact on GBM growth and has no toxicity in normal cells (astrocytes and microglia). Reaserches have further elucidated the potential mechanism of conventional antidepressants therapy for GBM.…”

The Organoid Era Permits the Development of New Applications to Study Glioblastoma

“…Leite and collaborators proved that 3D co-culture of human GB cell lines and microglia supports glioblastoma growth and migration creating a protective environment for GB [95]. They also displayed a new potential role of microglia in glioblastoma: microglia appear to modulate sensitivity to cytotoxic agents conferring drug resistance to the tumor.…”