Normal bronchial epithelial cell lines (BEAS-2B) and NSCLC cell lines (A549, GLC82, H1299, H1975, NIC-H157 and SW1573) were purchased from ATCC (Rockville, USA). All of these cells were cultured in RPMI-1640 complete medium containing 10% fetal bovine serum (FBS) (Thermo Scientific Hyclone, Utah, USA) and 1% penicillin/streptomycin (Thermo Scientific Hyclone, Utah, USA), and incubated at 37°C with 5% CO2 and saturated humidity. The medium was exchanged every 2–3 days. The cells were trypsinized with 0.25% trypsin (Thermo Scientific Hyclone, Utah, USA) when reaching 90% confluence.
Sw1573
Manufactured by American Type Culture Collection
Sourced in United States
The SW1573 is a laboratory equipment item designed for cell culture applications. It provides a controlled environment for the growth and maintenance of cell lines. The core function of the SW1573 is to regulate temperature, humidity, and gas composition to support optimal cell culture conditions.
Automatically generated - may contain errors
Lab products found in correlation
Nci h23, by American Type Culture Collection (7 mentions)
Nci h358, by American Type Culture Collection (7 mentions)
Calu 1, by American Type Culture Collection (7 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Nci h460, by American Type Culture Collection (5 mentions)
H1703, by American Type Culture Collection (4 mentions)
Sk lu 1, by American Type Culture Collection (4 mentions)
Beas 2b, by American Type Culture Collection (4 mentions)
Nci h1373, by American Type Culture Collection (4 mentions)
Nci h226, by American Type Culture Collection (4 mentions)
Nci h1299, by American Type Culture Collection (4 mentions)
H1299, by American Type Culture Collection (4 mentions)
Nci h2122, by American Type Culture Collection (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Hcc827, by American Type Culture Collection (4 mentions)
Calu 6, by American Type Culture Collection (4 mentions)
Nci h1792, by American Type Culture Collection (4 mentions)
H2122, by American Type Culture Collection (4 mentions)
36 protocols using sw1573
1
Cell Culture of Lung Epithelial Lines
2
Characterization of KRAS Mutant Cell Lines
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The KRAS mutant human cell lines HCC44, NCI-H23, NCI-H2122, SW1573 and NCI-H1373 were purchased from ATCC. H1792 is a gift from Professor Liu Xiangguo of Shandong University and HCC44-Luc is a gift from Professor Lu Weiyue of Fudan University. HCC44 were infected with luciferase retrovirus (HBLV-luc-BSD, LV57072301) to generate HCC44-Luc cells, and addition of appropriate concentrations of BSD to screen. THP-1 was obtained from Chinese Academy of Sciences. All cells were cultured at 37 °C in a 5% CO2 humidified incubator and grown in RPMI 1640 or DMEM medium containing 10% fetal bovine serum and penicillin/ streptomycin (100 U/ml, Life Technologies). All cell lines were tested for mycoplasma contamination.
Protein samples were subjected to SDS-PAGE and transferred to Polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford, MA). The membranes are incubated overnight at 4 °C in the appropriate primary antibody, then washed and incubated with the HRP-conjugated secondary antibody. Primary antibodies were as follows: TF (E9M6T, CST), ERK (4695, CST), P-ERK1/2 (4370, CST), AKT (P31749, Epitomics), P-AKT (Ser473) (ab81283, Abcam), CSF1(ab233387, Abcam), Bim (ab32158, Abcam) and GAPDH (AP0063, Bioworld).
Protein samples were subjected to SDS-PAGE and transferred to Polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford, MA). The membranes are incubated overnight at 4 °C in the appropriate primary antibody, then washed and incubated with the HRP-conjugated secondary antibody. Primary antibodies were as follows: TF (E9M6T, CST), ERK (4695, CST), P-ERK1/2 (4370, CST), AKT (P31749, Epitomics), P-AKT (Ser473) (ab81283, Abcam), CSF1(ab233387, Abcam), Bim (ab32158, Abcam) and GAPDH (AP0063, Bioworld).
3
Mesothelioma and Lung Cancer Cell Line Culture
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Mesothelioma (MERO-14, MERO-41, MERO-82, NO36, ONE58, MSTO-211H, NCI-H226, NCI-H2052) and lung cancer (NCI-H1703, -H211, -H526, -H522, -H520, -H460, -H358, -H1395, -H2122, -H2023, -H1651, A549, SW1573, DMS-114, DMS-53) cell lines were obtained from ATCC (Manassas, VA), DSMZ (Braunschweig, Germany), or Sigma-Aldrich (St. Louis, MO). Cells were cultured in the appropriate culture medium supplemented with 10% fetal bovine serum (FBS) at 37°C in humidified incubators under 5% CO2. Heparin sodium salt and growth factors were purchased from Sigma-Aldrich. GSK3052230 is formulated in 0.94 mg/mL sodium phosphate monobasic, 1.9 mg/mL sodium phosphate dibasic, 8.8 mg/mL sodium chloride, 0.2 mg/mL polysorbate 80, pH 7.0 at a stock concentration of 12.8 mg/mL. NVP-BGJ398 was purchased from Selleck Chemicals (Houston, TX) and dissolved in DMSO at a stock concentration of 20 mM.
4
Cell Culture and Compound Treatments
5
Evaluating KRAS-G12C Inhibitors in Cancer
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Three KRAS-G12C mutant lung adenocarcinoma, one colorectal and one pancreatic cancer cell line were involved in this study shown in Supplementary Table 1 . H358 and SW1573 were purchased from ATCC. PF97 and PF139 cells were established from malignant pleural effusion samples in cooperation with the West German Biobank Essen as described earlier [16 (link)]. The patients provided written informed consent and the experiments were approved by the Ethics Committee of the University Hospital Essen (#18-8208-BO).
Lonafarnib and tipifarnib (for in vitro experiments) were purchased from Sigma (St. Louis, MO, USA), while sotorasib, adagrasib and tipifarnib (for in vivo experiments) were obtained from Medchemexpress. For in vitro experiments, drugs were dissolved in dimethyl sulfoxide (DMSO) in 10 mM stock concentration and stored at −80 °C.
Lonafarnib and tipifarnib (for in vitro experiments) were purchased from Sigma (St. Louis, MO, USA), while sotorasib, adagrasib and tipifarnib (for in vivo experiments) were obtained from Medchemexpress. For in vitro experiments, drugs were dissolved in dimethyl sulfoxide (DMSO) in 10 mM stock concentration and stored at −80 °C.
6
Culturing SCLC and NSCLC Cell Lines
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SCLC cell lines H526 (CRL-5811), H446 (HTB-171), H249 (CRL-5827), H69 (HTB-119), H2171 (CRL-5929), H345 (HTB-180), H82 (HTB-175), H146 (HTB-173), H889 (CRL-5817) and H524 (CRL-5831), and NSCLC cell lines SW1573 (CRL-2170), H358 (CRL-5807), A549 (CCL-185EMT), H1838 (CRL-5899), H661 (HTB-183), H522 (CRL-5810), H1437 (CRL-5872), H2170 (CRL-5928), SW900 (HTB-59), H1993 (CRL-5909), SKLU-1 (HTB-57), H1703 (CRL-5889) and SKMES (HTB-58) were obtained from ATCC (Manassas, VA) and were cultured in RPMI 1640 medium (Gibco/BRL) supplemented with 10% (v/v) fetal bovine serum supplemented with L-glutamine and 1% (v/v) penicillin/streptomycin at 37 °C with 5% CO2.
7
Human NSCLC Cell Lines for Experimentation
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Human NSCLC cell lines A549, SK-LU-1, H1703, H358, H1993, H661, SW1573, H522, H226, H1437, H1838, H1975, H2170, and non-cancerous BEAS-2B were obtained from ATCC (Walkersville, MD, USA) and cultured in Roswell Park Memorial Institute complete medium (Cambrex, East Rutherford, NJ, USA) at 37°C in a humidified atmosphere of 5% CO2, 95% air, with passages 6–10 used for experimentation. Unless otherwise specified, reagents were obtained from Sigma (St. Louis, MO, USA). UK122 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Reagents for SDS-PAGE electrophoresis were purchased from Bio-Rad (Richmond, CA, USA) and Immobilon-P transfer membrane was purchased from Millipore (Millipore Corp., Bedford, MA, USA). Mouse anti-HABP2 antibody was purchased from Novus Biologicals (Littleton, CO, USA). Mouse anti-human nestin antibody that does not react with mouse or rat nestin (clone 10C2) was purchased from Millipore (Bedford, MA, USA). Mouse anti-β-actin antibody was purchased from Sigma (St. Louis, MO, USA). Secondary horseradish peroxidase-labeled antibodies were purchased from Amersham Biosciences (Piscataway, NJ, USA).
8
Characterization of Lung and Blood Cancer Cell Lines
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Human lung cancer cell lines (A549, NCIH1299, NCIH23, NCIH226, NCIH460, NCIH520, NCIH1703, SW1271, SW1573 and NCIH358) and blood cancer cell lines (KMS11 and RCH-ACV) were purchased from ATCC, JCRB and DSMZ. Antibodies were obtained from the following sources: NSD2 39879, H3K36me2 61019 and H3K36me2 39255 from Active Motif, NSD3 11345-1-AP from Proteintech, phospho ERK 4370 and H3K36me2 2901 from Cell Signaling, total ERK from BD Biosciences (anti-ERK1 554100 and anti-ERK2 610103), H3K36me2 ab9049 from Abcam, H3 07-690 and H3K27me3 07-449 from Merck Millipore and ACTB A5441 from Sigma Aldrich. Plasmid containing human NSD2 cDNA was purchased from Source BioScience (IMAGE ID: 100066394) and subcloned into pMSCV-FLAG-puro. The E1099K mutation was introduced using the QuikChange II XL Site-Directed Mutagenesis Kit (Agilent Technologies).
9
Non-Small Cell Lung Cancer Cell Line Cultivation
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The human NSCLC cell lines A549, Calu-1, Calu-6, NCI-H460, NCI-H292, NCI-H1299, NCI-H1730, NCI-H23, NCI-H3255, NCI-H520, NCI-H522, NCI-H596, HCC-827, SKMES-1, SW1573 and NCI-H1975 and the human epidermoid carcinoma A431 cells were purchased from ATCC (Manassas, VA), and cultured as recommended. 293H cells were purchased from Invitrogen (Carlsbad, CA). PC9, PC9GR4, PC9DR1 and HCC-827 GR5 were a kind gift of Dr. Pasi A. Jänne, Harvard University, Boston, MA [13 (link)]. PC9/ER, HCC-827/R1 and H3255/XLR cells are isogenic erlotinib- or XL-647-resistant lines derived from PC9, HCC-827 and NCI-H3255 cells, respectively, as previously described [53 (link)]. Additional information on cell cultures procedures is reported in the Supplemental Methods .
10
Cultivation of LUAD Cell Lines
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All LUAD cell lines (A549, Calu-1, H1792, H2030, MOR, and SW1573) used in this study were obtained from ATCC or Sigma-Aldrich (MOR only). The 634T cells were a generous gift from the laboratory of Kwok Wong (NYU). Cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 (Corning 10-040-CV) supplemented with penicillin–streptomycin and 10% fetal bovine serum. Spheroids were grown in medium supplemented with methylcellulose (Thermo Fisher Scientific M352-500). To prepare 100 mL of methylcellulose-containing medium, 1.2 g of methylcellulose was autoclaved in a bottle containing a stir bar. One hundred ml of medium was warmed to 37 °C and then added to the methylcellulose. The methylcellulose medium was then shaken vigorously and allowed to dissolve at 4 °C overnight, with stirring. Cells were maintained at 37 °C in a humidified incubator containing 5% CO2. All cell lines (and their derivatives) were confirmed to be mycoplasma-free using the MycoAlert mycoplasma detection kit (Lonza LT07-218) at the beginning and upon completion of all experiments.
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