Article Figures & Data
Figures
- Figure 1
Diagram of the construction patterns of different GBOs. The in vitro model of GBM was developed from the original 2-D and 3-D models and translated to the GBO model, and the genetically manipulated cerebral organoids, GLICO, PDO, and 3-D printed models were derived from the GBO model.
Tables
GBM model Cell resource Culture condition Characteristics Advantages and disadvantages References 2-D model Patient-derived GBM cells DMEM or DMEM/F12 culture medium with 10% FBS Monolayer adherent cells Easy cultivation, lack of stemness and differentiation ability 7,8 Sensitivity to therapy and radiation 10 3-D model Established cell line DMEM/F12 medium without serum, supplemented with EGF and bFGF Tumor balls Stemness, self-renewal, and differentiation ability; slow proliferation 7–9 Enhanced heterogeneity and drug resistance 11,12 GBO iPS cells DMEM/F12 containing B27, N2, and bFGF Brain tissue organoid Characteristics of human brain development 1,13 GBM cell line Matrix-free High stemness and strong cell-cell interaction 15 Genetically manipulated cerebral organoids iPS cells DMEM/F12 containing B27, N2, and bFGF Applied CRISPR/Cas9 technology Observation of the earliest steps of tumorigenesis in a human context with a defined genetic manipulation 17–19 Organoids derived from patients with c-Met mutation Accelerated differentiation into neurons for c-Met iPSC-A compared with control iPSC-A 20 PDO Patient tumor tissue DMEM/F12 containing B27, N2, and bFGF Organoids derived directly from glioblastoma specimens Generation of gradients of stem cell density and hypoxia with PDOs 22 DMEM/F12, Neurobasal medium, 1× GlutaMAX, NEAAs, penicillin–streptomycin, N2, and B27 Preservation of key characteristics and gene expression of the parental tumors Fast, recapitulation of heterogeneity and key features of glioblastomas 23–25 5% (w/v) GelMA and 0.25% (w/v) HA as the ECM Application of GelMA–HA hydrogels Maintenance of parental tumor features, such as the expression of key genes 26 DMEM, FBS, UltraGlutamine I, penicillin–streptomycin, and NEAA Description of specific steps High success rate and favorable preservation of patient heterogeneity 27,28 Analysis of genetic characteristics of parental patients and PDO models Mimicking of the tumor microenvironment and angiogenesis; high-throughput drug screening for precision medicine 29,30 GLICO mESCs and GSCs mESCs: DMEM-HG, KRS, MEM-NEAA, glutamine, β-mercaptoethanol, and LIF
GSCs: Neurobasal medium, B27, N2, bFGF, and EGFGBM spheroids in coculture with mouse embryonic stem cell (mESC)–derived early-stage cerebral organoids (eCOs) Favorable simulation of organoid compartments and infiltration patterns 31 GSCs and hESCs GSCs: Neurobasal medium, B27, N2, bFGF, and FGF
Cerebral organoids: DMEM/F12, Neurobasal medium, N2, B27, insulin, GlutaMAX, and MEM-NEAAGSCs co-cultured with cerebral organoids Simulation of invasion patterns and chemoradiotherapy resistance of patients with GBM 10,32 iPSCs and GSCs GSCs: Neurobasal medium, B27, L-Glutamine, Heparin, bFGF, and EGF
iPSCs: DMEM/F12 GlutaMAX, N2, B27, insulin, and L-glutamineCharacterization of GBM invasion into human brain at a quantitative or transcriptional level 33 3-D printed GBO GBM cells, ECM, and HUVECs GBM cells: DMEM, FBS, and penicillin–streptomycin
HUVECs: endothelial cell growth medium 2GBOs reconstructed from patient-derived glioma cells, vascular endothelial cells, and extracellular matrix, on the basis of 3-D printing technology Recapitulation of the structural, biochemical, and biophysical properties of native tumors; reproduction of clinically observed patient-specific resistance to treatment 37,38 GBM cells, monocytes, GASCs, and microglia GBM cells and monocytes: MEM, FBS, L-glutamine, and NEAA
GASCs: DMEM-HG medium
Microglia: microglia complete medium with serumUse of bioinks based on modified alginate to prepare tumor models incorporating tumor and stromal cells from glioblastoma Spatial organization of multiple cell types and recovery of protein and RNA at the single cell level 39 GSCs, macrophages, astrocytes, and neural stem cells GSCs: Neurobasal medium, B27, L-glutamine, bFGF, and FGF
Monocytes: RPMI 1640 medium, and FBS
Neural stem cells: complete NBM medium for GSCs
Astrocytes: astrocyte mediumModel comprising patient-derived GSCs, macrophages, astrocytes, and neural stem cells in HA-rich hydrogel Recapitulation of glioblastoma transcriptional profiles; promotion of hypoxic and invasive signatures; platform for drug response modeling 42,43 Application Targets Mechanisms References Biological function study Inflammation Dendritic polyglycerol sulfate limits GBM invasiveness by modulating microglial activation in GBOs. 45 Lipid metabolism GSCs have lower lipid droplet accumulation than non-GSCs in GBO models and xenograft tumors. 46 Invasion lncGRS antisense nucleotides decrease aggressive growth of tumors in GBOs. 47 Proliferation EGFRvIII mutation-induced astrogenesis and massive cell proliferation in human GBOs are observed. 48 Microenvironment Quiescent cells are partially responsible for tumor cell infiltration and invasion in GBOs. 49,50 Drug treatment Gamitrinib Gamitrinib inhibits cell proliferation, and induces cell apoptosis and death, in 17 primary glioma cell lines, 6 TMZ-resistant glioma cell lines, 4 neurospheres, and 3 PDOs. 56 UM-002 The novel BET inhibitor UM-002 decreases glioblastoma cell proliferation and invasion in GBOs. 57 Atorvastatin Atorvastatin has potent anti-angiogenic and apoptosis inducing effects against glioma spheroids. 58 Temozolomide Bioprinted GBOs reproduce clinically observed patient-specific resistance to treatment with concurrent chemoradiation and temozolomide. 38 Ruxolitinib Patients with GBM with than without LPS expression have higher levels of STAT1 and STAT2, and are more sensitive to ruxolitinib therapy. 59 Luteolin Luteolin decreases the proliferation of patient-derived glioma initiating cells and tumor organoids, but does not affect normal astrocytes. 60 Costunolide The TERT inhibitor costunolide effectively decreases cell viability in both primary GBO models and GBO models pre-treated with chemotherapy and radiotherapy. 61 MON MON decreases U87MG tumor size in GBOs and significantly decreases PARP expression. 62 GSC research PTEN GSCs are highly sensitive to proteasome inhibition, owing to an increased protein synthesis rate and loss of autophagy, associated with PTEN loss and activation of the PI3K/mTOR pathway. 68 Ciliogenesis Ciliogenesis-induced differentiation prevents the infiltration of GSCs in GBOs. 69 Phenotype Different microenvironments produced by transfer of GSC clonal populations from adherent culture to organoid conditions have different clonal phenotypes and are highly plasticized. 70 ZIKV ZIKV selectively eliminates GSCs from species-matched human mature cerebral organoids and GBM surgical specimens, but this effect is reversed by integrin αvβ5 inhibition. 71 ICB therapy HSPA7 promotes macrophage infiltration and SPP1 expression via upregulating YAP1 and LOX expression in GSCs. Knockdown of HSPA7 increases the efficiency of anti-PD1 therapy in the GBO model. 72
