Method Profile
Compare drug responses and transcriptomes in fresh-cell constructs and constructs remade after two-dimensional passage.
Assess how expansion changes patient-derived glioma pharmacology and model identity.
Nine analyzed glioma cases: 5 IDH-wild-type grade 4 glioblastomas, 3 IDH-mutant grade 2–3 astrocytomas and 1 oligodendroglioma.
Fresh P0 cells or two-dimensionally expanded P1/P2 cells were embedded in hyaluronan/gelatin constructs. Seven drug-free days were followed by 72 hours of treatment.
Matrix and cells
Patient-derived tumor constructs in defined hyaluronan/gelatin hydrogel. Fresh and previously expanded cells are distinguished.
Drugs and readouts
Temozolomide 100 μM, 500 μM or 1 mM; NSC59984 10 or 100 μM. ATP viability, live/dead imaging and RNA sequencing.
Compare constructs within each patient and cell-expansion state. Compare parent tumor, three-dimensional construct and two-dimensional culture transcriptomes.
Fresh versus expanded-cell pharmacology and transcriptional comparison.
29 biospecimens were procured; constructs were fabricated from 16 specimens, and 9 glioma cases were included in the reported analysis.
Untreated constructs; fresh-cell P0 constructs; constructs remade from P1/P2 cells; parent tumors and two-dimensional cultures.
Two-dimensional expansion produced gains or losses of temozolomide response. Parent tumors and three-dimensional constructs clustered by patient, whereas two-dimensional cultures converged on a mesenchymal expression class.
Hyaluronan-based patient-derived tumor constructs preserved patient-associated expression patterns better than two-dimensional cultures. Rebuilding constructs after cell expansion could either increase or reduce temozolomide sensitivity.
