Comprehensive Cancer Cell Line Characterization

All cancer cell lines (HT-29: colorectal adenocarcinoma, HCT-116: colorectal carcinoma, A549: lung epithelial carcinoma, PC3: prostate adenocarcinoma, Jurkat: acute T-cell leukemia, BC-1: primary effusion lymphoma) and the normal human embryonal lung fibroblast cell line MRC-5 were acquired from the American Type Culture Collection (ATCC). All cell lines were cultured as recommended by the suppliers. Culture media were purchased from Gibco (Thermo Fisher Scientific) and supplemented with 10 to 20% fetal bovine serum (HyClone, GE Healthcare Life Sciences) and 20–25 μg/mL gentamicin (Gibco, Thermo Fisher Scientific). All cells lines were thawed upon receipt from the distributor, amplified and subsequently batches were frozen in liquid nitrogen. Thawed lots were kept in culture no longer than 6 months.
Buffy coat preparations from healthy donors were obtained from the Blood Transfusion Center in Leuven, Belgium. Peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation over Lymphoprep™ (Axis Shield PoC AS; density 1.077 ± 0.001 g/mL) and cultured in cell culture medium (DMEM/F12, Gibco Thermo Fisher Scientific) containing 8% fetal bovine serum. Each donor consented to the use of his blood for research purposes.
Reference inhibitor compounds paclitaxel, vincristine and vinblastine were obtained from Selleckchem (Houston, Texas, USA) and stock solutions were prepared in DMSO at concentrations as specified in the manuscript.

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Herman J., Vanstreels E., Bardiot D., Prota A.E., Gaillard N., Gao L.J., Vercruysse T., Persoons L., Daems T., Waer M., Herdewijn P., Louat T., Steinmetz M.O., De Jonghe S., Sprangers B, & Daelemans D. (2024). 3-nitropyridine analogues as novel microtubule-targeting agents. PLOS ONE, 19(11), e0307153.

Publication 2024

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Variable analysis

independent variables

Cell lines (HT-29, HCT-116, A549, PC3, Jurkat, BC-1, MRC-5)
Reference inhibitor compounds (paclitaxel, vincristine, vinblastine)

dependent variables

Cell growth/proliferation

control variables

Culture media composition (DMEM/F12, 10-20% fetal bovine serum, 20-25 μg/mL gentamicin)
Culturing conditions (as recommended by suppliers)
Duration of cell line usage (thawed lots kept in culture no longer than 6 months)

controls

Positive controls: None specified
Negative controls: None specified

Annotations

Based on most similar protocols

Cell lines should be thawed upon receipt from the distributor, amplified, and subsequently batches frozen in liquid nitrogen. Thawed lots should be kept in culture no longer than 6 months (Input protocol).

Culture media should be supplemented with 10-20% fetal bovine serum and 20-25 μg/mL gentamicin (Input protocol).

Peripheral blood mononuclear cells (PBMC) should be isolated by density gradient centrifugation over Lymphoprep™ and cultured in DMEM/F12 medium containing 8% fetal bovine serum (Input protocol).

Reference inhibitor compounds (paclitaxel, vincristine, vinblastine) should be obtained from Selleckchem and stock solutions prepared in DMSO at specified concentrations (Input protocol).

For cytotoxicity evaluations in proliferating PBL cultures, nonadherent cells should be resuspended at 5 × 10^5 cells/mL in growth medium containing 2.5 μg/mL PHA. For cytotoxicity evaluations in resting PBL cultures, nonadherent cells should be resuspended at 5 × 10^5 cells/mL and treated for 72 h with the test compounds (Input protocol).

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