Three dimensional in vitro models of cancer: Bioprinting multilineage glioblastoma models

Abstract

Three dimensional (3D) bioprinting of multiple cell types within optimised extracellular matrices has the potential to more closely model the 3D environment of human physiology and disease than current alternatives. In this study, we used a multi-nozzle extrusion bioprinter to establish models of glioblastoma made up of cancer and stromal cells printed within matrices comprised of alginate modified with RGDS cell adhesion peptides, hyaluronic acid and collagen-1. Methods were developed using U87MG glioblastoma cells and MM6 monocyte/macrophages, whilst more disease relevant constructs contained glioblastoma stem cells (GSCs), co-printed with glioma associated stromal cells (GASCs) and microglia. Printing parameters were optimised to promote cell-cell interaction, avoiding the 'caging in' of cells due to overly dense cross-linking. Such printing had a negligible effect on cell viability, and cells retained robust metabolic activity and proliferation. Alginate gels allowed the rapid recovery of printed cell protein and RNA, and fluorescent reporters provided analysis of protein kinase activation at the single cell level within printed constructs. GSCs showed more resistance to chemotherapeutic drugs in 3D printed tumour constructs compared to 2D monolayer cultures, reflecting the clinical situation. In summary, a novel 3D bioprinting strategy is developed which allows control over the spatial organisation of tumour constructs for pre-clinical drug sensitivity testing and studies of the tumour microenvironment.

3D Bioprinted cancer models: Revolutionizing personalized cancer therapy

“…This was replicated by stimulating angiogenesis through cultured GSCs secreted factors present in the cell medium [ 32 , 35 ]. A recent study managed to recapitulate the heterogeneous glioblastoma tumor microenvironment in bioprinted constructs using an array of cancer and stromal cells including MM6 monocyte/macrophages, U87MG glioblastoma cells, glioblastoma stem cells, microglia and glioma associated stromal cells (GASCs) providing a more realistic chemotherapeutic response [151] ( Fig. 4 -C).…”

Biomaterials and 3D Bioprinting Strategies to Model Glioblastoma and the Blood–Brain Barrier

“…Another multinozzle extrusion‐based bioprinting method was used to produce a 3D model with GBM cell lines or GSCs, patient‐derived GBM‐associated stromal cells (GASCs), and microglia in an alginate‐based hydrogel. [ 188 ] The alginate was functionalized with RGDs, and in some groups with HA and collagen. The 3D multicellular models showed moderately enhanced resistance to TMZ and enhanced resistance to cisplatin, a compound that failed many clinical trials but showed promising results in 2D cultures, indicating potential application of this system for more reliable preclinical drug efficacy evaluations.…”

Fabrication techniques of biomimetic scaffolds in three‐dimensional cell culture: A review

“…Recently, bioprinted alginate–collagen–hylauronic acid matrix comprising of glioblastoma cancer cells was fabricated to control the spatial organization of tumor constructs applied for preclinical drug sensitivity and to study the tumor microenvironment (Hermida et al, 2020). The study shows that the 3D bioprinting of cells has negligible effect on cell viability, and more resistance to anticancer drugs (resembling in vivo condition) than 2D culture.…”