Rapid 3D Bioprinting of Glioblastoma Model Mimicking Native Biophysical Heterogeneity

Abstract

Glioblastoma multiforme (GBM) is the most lethal primary brain tumor characterized by high cellular and molecular heterogeneity, hypervascularization, and innate drug resistance. Cellular components and extracellular matrix (ECM) are the two primary sources of heterogeneity in GBM. Here, biomimetic tri‐regional GBM models with tumor regions, acellular ECM regions, and an endothelial region with regional stiffnesses patterned corresponding to the GBM stroma, pathological or normal brain parenchyma, and brain capillaries, are developed. Patient‐derived GBM cells, human endothelial cells, and hyaluronic acid derivatives are used to generate a species‐matched and biochemically relevant microenvironment. This in vitro study demonstrates that biophysical cues are involved in various tumor cell behaviors and angiogenic potentials and promote different molecular subtypes of GBM. The stiff models are enriched in the mesenchymal subtype, exhibit diffuse invasion of tumor cells, and induce protruding angiogenesis and higher drug resistance to temozolomide. Meanwhile, the soft models demonstrate enrichment in the classical subtype and support expansive cell growth. The three‐dimensional bioprinting technology utilized in this study enables rapid, flexible, and reproducible patient‐specific GBM modeling with biophysical heterogeneity that can be employed by future studies as a tunable system to interrogate GBM disease mechanisms and screen drug compounds.

Microengineered perfusable 3D-bioprinted glioblastoma model for in vivo mimicry of tumor microenvironment

“…We also show that expressed genes in our fibrin 3D-bioink provides essential cues to enhance tumor growth and survival; however, they are inefficiently expressed in standard 2D culture. Up-regulated gene expression levels in our system were also seen in similar GB 3D models (17,45,46); however, these models lack the whole complexity of GB TME (astrocytes, microglia, endothelial cells, and pericytes) together with the GB cells, as presented in this manuscript. This further emphasized that our 3D-bioink can serve not only as a drug Fig.…”

3D bioprinting of complex tissues in vitro: state-of-the-art and future perspectives

“…Cancer remains a significant public health issue worldwide due to its high occurrence, mortality, and economic impact. 3D bioprinting offers an opportunity to advance the in vitro modeling of various cancers types, such as brain cancer (Tang et al 2020 , 2021 ), pancreatic cancer (Hakobyan et al 2020 ), liver cancer (Ma et al 2018a , b ), lung cancer, colorectal cancer, ovarian cancer (Xu et al 2011 ), breast cancer (Hribar et al 2015 ; Zhu et al 2016 ), and metastatic models (Meng et al 2019 ; Zhu et al 2016 ), owing to its ability to recapitulate the complex cellular and material heterogeneities.…”

Glioblastoma Vasculature: From its Critical Role in Tumor Survival to Relevant in Vitro Modelling

“…Printed cells usually include cancer cell lines or patientderived tumor cells, ECs, fibroblasts or immune cells, while alginate, gelatin, polyethyleneglycol (PEG) methacrylate or decellularized ECM comprise the models' matrix. These models are usually very reproducible and adjustable since their production is entirely dependent on computer programs and no human manipulation is directly involved (Tang et al, 2021). In contrast with 2D cell cultures, 3D-bioprinted models maintain in vivo stemness properties, gene expression and vascularization tendency.…”

“…Recently, 3D bioprinting has revealed its applicability and usefulness in the neuro-oncology field, specifically in the assembly of GBM in vitro models to study neovascularization. Both GBM cells and ECs bioprinted within ECM-like materials sprout, forming tubular structures (Figure 4) (Table 1) (Han et al, 2020;Tang et al, 2021). Moreover, pro-angiogenic genes expression and cells' sprouting is regulated by matrix biophysical properties, such as stiffness (Tang et al, 2021).…”

“…Both GBM cells and ECs bioprinted within ECM-like materials sprout, forming tubular structures (Figure 4) (Table 1) (Han et al, 2020;Tang et al, 2021). Moreover, pro-angiogenic genes expression and cells' sprouting is regulated by matrix biophysical properties, such as stiffness (Tang et al, 2021). Therefore, these models mimic in vivo angiogenesis and GSCs transdifferentiation.…”