H2110, H1915, H650, H1623, H2126, HCC827, A549, H810, H1048, H1355 and HEK293T, and Beas‐2B were purchased from ATCC. The H157 cell line was a kind gift from Dr Phillip Dennis (Johns Hopkins University). All cell lines were maintained in RPMI1640 media (Invitrogen) supplemented with 2 mM L‐glutamine (Gibco), 10% FBS (Lonza), and 1% Pen/Strep (Gibco). 293FT cells (Invitrogen) were maintained in DMEM media (Gibco) supplemented with 10% FBS (Lonza) 4 mM L‐glutamine (Gibco), 1 nM sodium pyruvate (Gibco), and 0.1 mM NEAA (Gibco). All experiments were performed within 6 months of cell line delivery and cell lines were authenticated by DNA sequencing at the start of experiment. After 6 months, cells were discarded and new cells ordered from ATCC. Mycoplasma testing was performed on regular basis.
H1048
Manufactured by American Type Culture Collection
Sourced in United States
H1048 is a Biological Safety Cabinet that provides protection for personnel, product, and the environment when handling hazardous biological materials. It features a stainless-steel work surface, HEPA filtration, and adjustable airflow controls to maintain a safe work environment.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 media, by Thermo Fisher Scientific (3 mentions)
Hcc827, by American Type Culture Collection (2 mentions)
Hek293t, by American Type Culture Collection (2 mentions)
Dms53, by American Type Culture Collection (2 mentions)
Sbc 3, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Penicillin streptomycin, by Fujifilm (2 mentions)
Rpmi 1640, by Fujifilm (2 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
H1355, by American Type Culture Collection (1 mentions)
H2110, by American Type Culture Collection (1 mentions)
H1915, by American Type Culture Collection (1 mentions)
293ft cell, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Dmem media, by Thermo Fisher Scientific (1 mentions)
10 protocols using h1048
1
Cell Lines Maintenance Protocol
2
Small Cell Lung Cancer Cell Lines: Characterization and Maintenance
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The human SCLC cell lines SBC‐3 and SBC‐5, were kindly provided by Dr. K. Kiura (Okayama University, Okayama, Japan). DMS237 and DMS273‐G3H were obtained as reported previously.21 Briefly, DMS273‐G3H was established using tumor cells from a bone metastasis after we had implanted DMS273 orthotopically into the left lung of nude mice. H196 and H1048 were purchased from ATCC (Manassas, VA, USA). DMS114 and DMS454 were purchased from the European Collection of Authenticated Cell Cultures (Porton Down, Hampshire, UK). All cells were maintained in RPMI‐1640 medium supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (10 μg/mL) in a humidified, 5% CO2 incubator at 37°C. All cells were passaged for a total of less than 3 months before being replaced by frozen early‐passage stocks. Cells were regularly screened for mycoplasma using a MycoAlert Mycoplasma Detection Kit (Lonza, Rockland, ME, USA). Golvatinib was obtained from Selleck Chemicals (Houston, TX, USA), and crizotinib was obtained from Active Biochem (Kowloon, Hong Kong).
3
Small Cell Lung Cancer Cell Culture and Compound Preparation
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H1048 (SCLC-P), H211 (SCLC-P), H146 (SCLC-A), H526 (SCLC-P), H345 (SCLC-A), and H209 (SCLC-A) cells were purchased from ATCC (Manassas, VA, USA). Cells were cultured in RPMI 1640 media (Thermo Fisher, 11875093, Waltham, MA, USA), supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Sigma, St. Louis, MO, USA, 20K514), and 100 µg/mL penicillin G/streptomycin at 37 °C in a humified, 5% CO2 incubator. ABT-199 (MedChemExpress, HY-15531, Monmouth Junction, NJ, USA, and LC Laboratories, V-3579, Woburn, MA, USA) and Ganetespib (MedChemExpress, HY-15205, Monmouth Junction, NJ, USA) were dissolved in dimethyl sulfoxide (DMSO), and stable drugs were stored at −20 °C in the dark. The final concentration of DMSO was 0.1%.
4
SCLC Cell Line Cultivation Protocol
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Human SCLC cell lines H446, DMS79, H524, H526, H82, H847, H841, H196, H211, H146, SHP77, H345, H2196, H1436, H865, H1522, H1341, H1105, H1048, H1092, H1876, H69, DMS53, and H2029 were obtained from ATCC (Manassas, VA, USA) or Sigma-Aldrich (St. Louis, MO, USA). The human patient-derived xenograft (PDX) cell line NJH29 was generously provided by Dr. Julien Sage (Stanford University, Stanford, CA, USA). For experimentation, cell lines were cultured in RPMI-1640 supplemented with 10% FBS, 100 IU/mL penicillin, and 100 μg/mL streptomycin. Cells were maintained in a 37 °C humidified chamber with 5% CO2. All cell lines used were passaged for less than 6 months and regularly tested for Mycoplasma contamination using a MycoAlert Plus Kit (Lonza).
5
Cell Line Establishment and Culture
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Cell Culture. H446, H187, H460, H1048, H1436, Calu-6, MDA-MB-468, and HEK293T cells were purchased from the American Type Culture Collection, SCLC-21H was from the German Collection of Microorganisms and Cell Cultures, and COR-L279 and COR-L88 were from Sigma.
LX95 SCLC line was established by growing the single cells isolated from the LX95 PDX tumors in HITES medium (DMEM:F12 medium supplemented with 0.005mg/ml insulin, 0.01 mg/ml transferrin, 30 nM sodium selenite, 10 nM hydrocortisone, 10 nM beta-estradiol, 1x Glutamax, 5% heat-inactivated fetal bovine serum, 1x penicillin/streptomycin). H446 cells with ectopic expression of BCL2 were established by transfection of H446 cells with Flag-BCL2 (a gift from Clark Distelhorst (17) ; Addgene plasmid #18003) followed by selection with 0.4 mg/ml G418 (Roche). All cell lines were grown in culture media recommended by the suppliers and were maintained in humidified incubators at 37 o C. MDA-MB-468 was grown at 100% air, while all other lines were at 5% CO2. All cell lines tested negative for mycoplasma, and short tandem repeat analysis was performed to authenticate commercial cell lines.
LX95 SCLC line was established by growing the single cells isolated from the LX95 PDX tumors in HITES medium (DMEM:F12 medium supplemented with 0.005mg/ml insulin, 0.01 mg/ml transferrin, 30 nM sodium selenite, 10 nM hydrocortisone, 10 nM beta-estradiol, 1x Glutamax, 5% heat-inactivated fetal bovine serum, 1x penicillin/streptomycin). H446 cells with ectopic expression of BCL2 were established by transfection of H446 cells with Flag-BCL2 (a gift from Clark Distelhorst (17) ; Addgene plasmid #18003) followed by selection with 0.4 mg/ml G418 (Roche). All cell lines were grown in culture media recommended by the suppliers and were maintained in humidified incubators at 37 o C. MDA-MB-468 was grown at 100% air, while all other lines were at 5% CO2. All cell lines tested negative for mycoplasma, and short tandem repeat analysis was performed to authenticate commercial cell lines.
6
Small Cell Lung Cancer Cell Line Cultures
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We used 12 SCLC cell lines: SBC‐3 and SBC‐5 from the Japanese Collection of Research Bioresources Cell Bank, H69, H69AR, H719, H1048, H1105, H1417, DMS53, and H1882 from the American Type Culture Collection, and MS‐1 and Lu‐139 from the Riken Cell Bank. SBC‐3 and SBC‐5 were cultured in MEM‐EAGLE medium (Sigma‐Aldrich) with 10% fetal bovine serum (FBS; Biowest) and 1% penicillin/streptomycin (Fujifilm Wako). H69AR was maintained in RPMI‐1640 (Fujifilm) with 20% FBS and 1% penicillin/streptomycin. The other SCLC cell lines were maintained in RPMI‐1640 with 10% FBS and 1% penicillin/streptomycin. These cell lines were obtained between 2008 and 2017 and were routinely examined for the absence of mycoplasma.
7
Cell Line Maintenance Protocols
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The H446 (HTB-171) and H1048 (CRL-5853) SCLC cell lines were purchased from the American Type Culture Collection (ATCC) and were maintained in ATCC recommended culture medium. HUVECs and HEK293T cells were cultured in DMEM (Corning) containing 10% FBS (Gibco) and 1% penicillin and streptomycin (Gibco).
8
Overexpression of miR-141 in SCLC
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SCLC cell lines (H446 and H1048) obtained from the American Type Culture Collection (ATCC) were maintained in the recommended CM in an incubator containing 5% CO2 at 37 °C. Dulbecco’s modified Eagle’s medium (DMEM, Corning) containing 10% FBS (Gibco) and 1% penicillin and streptomycin (Gibco) was used to culture HUVECs and EAhy.926 cells. To develop SCLC sublines that stably overexpressed miR-141, SCLC cells were screened with puromycin after lentivirus infection for half a month and then maintained in CM with low-dose puromycin.
9
Lung Cancer Cell Lines Sensitivity to BIRC5 Inhibitor
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Human SCLC cell lines SBC3 and SBC5 were purchased from the Japanese Collection of Research Bioresources Cell Bank. SCLC cell line H1048, non-small cell lung cancer (NSCLC) cell lines A549, EBC1, and EREF-LC-KJ, and human lung fibroblasts (HLFs) were purchased from American Type Culture Collection. All cells were cultured with RPMI-1640 (FUJIFILM Wako Pure Chemical Corporation) with 10% fetal bovine serum (Biosera). BIRC5 inhibitor (YM155) was purchased from Selleck Chemicals and used to culture SBC3, SBC5, and H1048 cells at concentrations of 2 nM, 3 nM, and 7 nM for 48 h to inhibit the function of BIRC5.
10
Establishing Gefitinib-Resistant NSCLC Cell Lines
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Pulmonary epithelial BEAS-2B cells and NSCLC (HCC827, H1870, H1048, PC9 and A459) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Gefitinib-resistant cell lines PC9/R and A459/R were obtained by gradually increasing the gefitinib concentration. All cells were cultured in the 1640 medium (HyClone, Logan, UT, USA) and incubated in the 5% CO2 incubator at 37°C. The log-phase cells were harvested.
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