Method Profile
Compare established MCF-7 spheroids in EGF-loaded and unloaded alginate cryogels for three-day drug response and associated cellular phenotypes.
Assess how matrix-associated growth-factor presentation changes measured drug sensitivity in a controlled cell-line setting.
MCF-7 human breast-cancer cells in porous 2% calcium-crosslinked alginate cryogels. EGF-loaded and unloaded matrices are compared, with a separate two-dimensional soluble-EGF profiling condition.
The Results select a nominal 600 ng/mL EGF condition. About 50% of matrix-associated EGF is released within 48 hours, with about 40% retained at day 7. Drug response is measured after three days; loading is distinct from constant measured soluble exposure.
Drugs and viability
Doxorubicin, epirubicin, 5-fluorouracil and paclitaxel; metabolic viability normalized to the corresponding no-drug condition.
Phenotype and profiling
Spheroid area, Ki-67, Transwell migration after dissociation, annexin-V/PI, TUNEL/caspase-3, EMT-associated markers, RNA sequencing, cytokine ELISA and metabolomics.
Keep cell-line source, area versus volume, transcript versus protein and association versus causal mechanism distinct. The paclitaxel IC50 in EGF-loaded cultures remains unestimated.
One established MCF-7 cell line; quantitative measurements use three replicate samples.
EGF-loaded versus unloaded alginate cryogels; separate two-dimensional soluble-EGF transcriptome; matched untreated drug controls.
EGF loading increased doxorubicin IC50 from 1.93 ± 0.10 to 4.53 ± 0.52 μM, epirubicin from 0.76 ± 0.08 to 2.64 ± 0.35 μM, and 5-fluorouracil from 9.23 ± 0.94 to 13.94 ± 1.39 μM. Paclitaxel control IC50 was 2.32 ± 0.40 μM; the loaded-group IC50 was not estimable because of insufficient cell loss. Ki-67 increased and the apoptotic fraction decreased.
Primary study comparisons of the named models, exposures and measured outcomes.
MCF-7 cells grown in EGF-loaded alginate cryogels showed increased proliferation and reduced sensitivity to several drugs. The study links those changes to matrix and growth-factor signaling, with gene-expression and metabolite associations supplying hypotheses rather than patient-response validation.
