Chapter 1 of 5 · Fundamentals
Platform rationale
Introduction and rationale for patient-derived GBM tissue platform
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What the experts said
Traditional methods like animal models and 2D cell cultures fall short in replicating the complexity of glioblastoma microenvironment.
Patient GBM tissue is the best option for closely mimicking the microenvironment of GBM and testing therapeutic interventions.
The microfluidic chip can maintain patient-derived GBM tissue for 8 days and in some cases 12 days in the laboratory.
Fresh GBM tissue is obtained during surgery with patient consent, placed in nutrient media immediately, and transferred to the lab within one hour.
The microfluidic chip uses a cylindrical shape with an inner tube connected to a syringe containing nutrient media with or without treatment.
The treatments tested were temozolomide (the classic treatment) plus protein arginine methylation inhibitors.
The chip is connected to an effluent tube collecting released cytokines and placed in a Harvard syringe pump with a flow rate of 3 microliters per minute over 8 to 12 days.
Effluent containing cytokines is collected every 24 hours and stored at -80°C to maintain cytokine integrity.
The R&D Systems protein profiler membrane allows 105 human cytokines to be tested on a single membrane in duplicate form.
Selected cytokines from the 105-cytokine panel were validated by ELISA.
The study includes data from 13 patients over 12 days.
H&E staining shows GBM tissue cells maintain their integrity after 8 days on the chip and also at 12 days.
GBM tissue viability in the microfluidic chip shows minimal difference over the first 4 days (96 hours).
After 96 hours (4 days), GBM tissue begins to decrease and change patterns, with more pronounced changes at 12 days.
EGF and MMP9 are significantly reduced under the novel treatment (temozolomide plus protein arginine methylation inhibitors) compared to control.
SENES3-like 1, a cytokine mainly related to colon cancer, is abundant in GBM and increases against treatment, potentially indicating a resistance mechanism.
The microfluidic platform demonstrates viability of maintaining glioblastoma tissue for 8–12 days while testing different treatments against control.
The microfluidic model represents a potential GBM model for testing therapeutic interventions and observing effects over 8 to 12 days.
