Effects of three-dimensional collagen scaffolds on the expression profiles and biological functions of glioma cells

Abstract

Three-dimensional (3D) culture has been increasingly used to investigate tumor cell biology for improved simulation of the natural developing environment. However, the way in which 3D culture affects the gene expression and biological functions of glioma cells remains to be fully elucidated. In the present study, 3D culture environments were established using collagen scaffolds with different pore sizes, followed by the comparison of gene expression profiles and associated biological functions of glioma cells, including the U87, U251 and HS683 cell lines, in 3D collagen scaffolds with conventional two-dimensional (2D) cultured cells. Finally, the possible signaling pathways regulating these differences were investigated. It was found that the 3D collagen scaffold culture upregulated the expression of genes associated with stemness, cell cycle, apoptosis, epithelia-mesenchymal transition, migration, invasion and glioma malignancy, and induced the corresponding functional changes. Apoptotic pathways, the Wnt pathway, Sonic Hedgehog pathway and Notch pathway, may be involved in the regulation of these changes. The aperture size of the collagen-scaffold did not appear to affect the gene expression or functions of the glioma cells. The results of the study suggested that the 3D collagen scaffold enhanced the malignancy of glioma cells and may be a promising in vitro platform for investigations of glioma.

Involvement of NOS2 Activity on Human Glioma Cell Growth, Clonogenic Potential, and Neurosphere Generation

“…Allen et al reported the origin of the U-87 MG cell line as mislabeled and unknown; they compared the original U-87MG cell line firstly identified at Uppsala University with the commercial U-87MG cell line from ATCC HTB 14TM. Genotyping analysis revealed that the ATCC and Uppsala U-87 MG cell lines were from distinct origins [ 57 ]; however, other recent studies [ 58 , 59 , 60 ] have demonstrated that the U-87MG ATCC cell line has several typical characteristics of glioblastoma through analyses of cell morphology and gene expression profile, even though the origin patient is unknown. Thus, U-87 MG can be used as a bonafide human glioblastoma cell line.…”

Transcriptomic Profiling of 3D Glioblastoma Tumoroids for the Identification of Mechanisms Involved in Anticancer Drug Resistance

“…Tejero, et al found that the 3D glioblastoma organoids promoted the appearance of a quiescent cell population that represented the self-renewal ability high therapy resistance, and mesenchymal gene signatures (13). Furthermore, Jia, et al found that the expression of genes associated with stemness, cell cycle, apoptosis, epithelia-mesenchymal transition, migration, invasion, and glioma malignancy, were upregulated in 3D glioma cell culture (14). Despite the number of 3D glioblastoma studies, the transcriptomic profile and molecular pathway need to be further explored in order to understand the global picture of the underlying drug resistance mechanism.…”

Methods for in vitro modeling of glioma invasion: Choosing tools to meet the need

“…Scaffoldings are 3D environments that bridge the gap between conventional 2D assays that are cheap and reproducible yet do not comprehensively model the brain physiology and animal models, which introduce technical difficulties, high costs, and low reproducibility (Rao et al, 2014). Moreover, 3D cultured GBM cells behave differently than their 2D monolayer counterparts, more closely resembling in vivo phenotypes (Jia et al, 2018;Lv et al, 2016). Stemnessassociated gene and protein expression levels, invasive capacity, glioma malignancy, and therapeutic resistance all increase in tumor cells cultured with 3D scaffold systems compared to 2D assays (Dai, Ma, Lan, & Xu, 2016;Goncalves et al, 2017;Jia et al, 2018;Jiguet Jiglaire et al, 2014;Lv et al, 2016;.…”

“…Migration using a scratch assay is monitored at distinct intervals capturing different positions along the scratch and measuring the distance between both wound edges (Figure 5a, left). Distances measured on latter images are normalized to the same point immediately following the scratch, generating a distance closed during the experiment duration (Figure 5a, middle;Arslan et al, 2007;Jia et al, 2018;Lin, Kuo, Chou, & Lu, 2008). Alternatively, cells within the scratch area can be counted (Figure 5a, right;Gessler et al, 2010;Priester et al, 2013;Wei, Ryu, Choi, & McLarnon, 2008).…”

“….In order to mimic microenvironmental and microanatomic features found in the brain, both natural and synthetic materials have been utilized for scaffold production including hyaluronic acid (HA), collagens, Matrigel, astrocyte-rich stroma, chitosan, alginate, reconstituted basement membrane, poly-caprolactone, polyacrylamide hydrogels, and composite hydrogels(Table 1)(Agudelo-Garcia et al, 2011;Erickson et al, 2018;Goncalves et al, 2017;P. Gritsenko et al, 2017;Jia et al, 2018;Rao et al, 2014;Yang, Motte, & Kaufman, 2010). Glioma cells can be embedded and cultured as single cells or neurospheres in 3D hydrogel systems(Figure 3a; J. M Heffernan et al, 2015;Jiguet Jiglaire et al, 2014;Lv et al, 2016)…”