NSCLC Cell Line Characterization and CSC Propagation

Human NSCLC cell lines A549 (KCLB No. 10185), NCI-H1299 (KCLB No. 25803), NCI-H1650 (KCLB No. 91650), and HCC827 (KCLB No. 70827) were obtained from the Korean Cell Line Bank (Seoul, Republic of Korea). The identity of each cell line was confirmed by STR profiling: A549 (D3S1358: 16; vWA: 14; FGA: 23; Amelogenin: X,Y; TH01: 8,9.3; TPOX: 8,11; CSF1PO: 10,12; D5S818: 11; D13S317: 11; D7S820: 8,11), NCI-H1299 (D3S1358: 17; vWA: 16,18; FGA: 20; Amelogenin: X; TH01: 6,9.3; TPOX: 8; CSF1PO: 12; D5S818: 11; D13S317: 12; D7S820: 10), NCI-H1650 (D3S1358: 18; vWA: 18; FGA: 20,23.2; Amelogenin: X; TH01: 9.3; TPOX: 11; CSF1PO: 11; D5S818: 11; D13S317: 11; D7S820: 8,9), and HCC827 (D3S1358: OL; vWA: 18; FGA: 22,24; Amelogenin: X; TH01: 6; TPOX: 8; CSF1PO: 11; D5S818: 12; D13S317: 9; D7S820: 11,12). CSCs were propagated from NSCLC cell lines as tumorspheres under CSC-selective culture conditions, and their stem properties were confirmed as previously described [23 (link),24 (link),25 (link)]. NSCLC tumorsphere cells were cultured in serum-free DMEM/F12 containing 20 ng/mL EGF, 20 ng/mL bFGF, 1 × B-27, 5 μg/mL heparin, 2 mM L-glutamine, and 1% penicillin/streptomycin. Tumorspheres were subcultured through dissociation with Accutase. The cells were maintained at 37 °C in a humidified CO₂ incubator with 5% CO₂ (Thermo Scientific, Vantaa, Finland).

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Han J.M., Kim S.M., Kim H.L., Cho H.J, & Jung H.J. (2023). Natural Cyclophilin A Inhibitors Suppress the Growth of Cancer Stem Cells in Non-Small Cell Lung Cancer by Disrupting Crosstalk between CypA/CD147 and EGFR. International Journal of Molecular Sciences, 24(11), 9437.

Publication 2023

NCI-H1299 NCI-H1650 HCC827

Accutase Amelogenin Cell lines Cells Glutamine Heparin Human Korean Nsclc Penicillin Serum Stem Streptomycin

Corresponding Organization : Sun Moon University

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Variable analysis

independent variables

CSC-selective culture conditions

dependent variables

Stem properties of NSCLC tumorsphere cells

control variables

Serum-free DMEM/F12 medium
20 ng/mL EGF
20 ng/mL bFGF
1 × B-27
5 μg/mL heparin
2 mM L-glutamine
1% penicillin/streptomycin
37 °C temperature
5% CO2

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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