Fig. 2: Tunable model of the human invasive TME enables multiplexed analysis of glioma markers and comparison to in vivo marker expression.

a Diagram of brain TME model setup using patient-derived GSCs, human astrocytes, and human microglia in a hyaluronan-based matrix and 96-well tissue culture insert format. b Representative flow cytometry plots showing the ability to distinguish astrocyte, microglia, and glioma cell populations and determine glioma-specific proliferation (Ki67⁺), stemness (CD71⁺), and cell death. Isotype controls are shown in gray. Also shown is a representative fluorescence image of the porous membrane for invasion quantification. Scale bar is 50 µm. c Representative fluorescence image within the gel with glioma cells (blue), astrocytes (green), and microglia (magenta). Scale bar is 50 µm. d Quantification of cell viability under different media formulations for up to three days in hydrogel culture. e Quantification of cell viability following different enzymatic gel degradation protocols. f, g Comparison of %Ki67⁺ cells (f) and %CD71⁺ cells (g) for three in vitro cancer models and in vivo xenograft implants. In vitro data obtained by flow cytometry; in vivo data obtained from tissue sections, with the tumor border visually demarcated based on nuclear staining. Legends in f, g are the same. Comparisons conducted by unpaired t-tests, *p < 0.05 and **p < 0.01 for n = 3–4.