Method Profile
Maintain precision-cut peritoneal-metastasis tissue in tuned hyaluronan hydrogels and compare drug response with and without patient ascites.
Investigate how native tumor tissue and its fluid environment affect short-term drug response.
Resected peritoneal-metastasis slices, 300 μm thick, from a 21-patient collection with distinct subsets per assay. A 2% hyaluronan (HA)-RGD-PQ hydrogel supports the mixed native tissue.
Optimized 2% hyaluronan with a storage modulus around 350 Pa; preservation measured through 12 days. The drug assay uses two days of culture, one day of serum starvation, 24-hour pretreatment with 5% ascites and 48-hour drug exposure, with pre/post-treatment resazurin measurements.
Preservation comparisons
PTFE, Matrigel, two- versus three-dimensional hyaluronan, stiffness/degradability-matched hyaluronan and PEG, plus RGD/RDG controls.
Drug and context measurements
Cisplatin, doxorubicin, erlotinib and TM5441 dose-response; resazurin metabolic change, live/dead staining, histology, cell-state RNA profiles and ascites PAI-1.
Do not substitute area preservation for viability or tissue-wide metabolic inhibition for a tumor-specific cell count. Separate preservation subsets, three-patient profiling, two-patient ascites profiling and six-fluid TM5441 comparison.
Fresh samples from 21 patients; main preservation experiments use at least two patients with at least three technical replicates where specified. Day-8 tissue profiling uses three patients and ascites profiling two patients. The TM5441 comparison uses six ascites samples.
PTFE/Matrigel, material-property controls, serum versus ascites, different fluid/tumor pairings and untreated drug controls.
Optimized hyaluronan supported tissue preservation; stiffer gels could retain area with poorer viability. Ascites did not improve basic preservation but increased drug tolerance. TM5441 response was associated with PAI-1, but PC294/PC307 fluids did not sensitize NCC-MC-308/NCC-MC-375 tumors.
Primary study comparisons of the named models, exposures and measured outcomes.
Optimized hyaluronan hydrogels maintained peritoneal-metastasis explants while limiting tissue contraction. Adding ascites changed drug sensitivity without improving basic tissue preservation, and PAI-1 inhibitor responses depended on the fluid sample as well as the tumor.
