H1975 (CRL‐5908, ATCC, LCG standards, Wesel, Germany), HCC827 (CRL‐2868, ATCC, LCG standards, Wesel, Germany) and A549 (CCL‐185, ATCC, LCG standards, Wesel, Germany) cells were grown in RPMI medium containing 10% fetal calf serum and 1% penicillin‐streptomycin (Gibco, Thermo Fischer Scientific, Waltham, MA, USA). The cells were cultivated at 37 °C in 5% CO2.
Crl 2868
Manufactured by American Type Culture Collection
Sourced in Germany, United States
The CRL-2868 is a cell line obtained from Homo sapiens (human). It is a fibroblast cell line derived from human skin. The core function of this cell line is to serve as a research tool for various scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Ccl 185, by American Type Culture Collection (2 mentions)
Crl 5908, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Hcc827, by American Type Culture Collection (1 mentions)
Met 5a cells, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Erlotinib osi 744, by Selleck Chemicals (1 mentions)
Crl 5935, by American Type Culture Collection (1 mentions)
4 protocols using crl 2868
1
Cell Culture of NSCLC Lines
2
Mesothelial Cell Co-Culture with Lung Cancer Cells
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Pleural mesothelial MeT-5A cells (CRL-9444™) were obtained from the American Type Culture Collection (American Type Culture Collection (ATCC), Manassas, USA) and were cultured in Medium 199 containing 1.5 g/L sodium bicarbonate, 10% fetal bovine serum (FBS), epidermal growth factor (3.3 nM), hydrocortisone (400 nM), bovine insulin (870 nM), HEPES (20 mM) and trace elements (H2SeO3, MnCl2, Na2SiO3, (NH4)6Mo7O24, NH4VO3, NiSO4 and SnCl2). A549 (CCL-185™) and CL1-5 cell lines were cultured in F-12K Medium and RPMI-1640 medium supplied with 10% FBS, respectively. Epidermal growth factor receptor (EGFR) mutation lung cancer cell lines NCI-H1975 (H1975, CRL-5908™) and HCC827 (CRL-2868™) were obtained from ATCC and cultured in RPMI-1640 medium supplied with 10% FBS. MeT-5A cells were co-cultured with various lung cancer cell lines using a transwell culture system (pore size, 1 µm; Corning Incorporated, Corning, NY, USA) for 24 h and 48 h. Lung cancer cells were grown in the upper chamber with MeT-5A cells in the bottom chamber. MeT-5A cells cultured alone served as controls.
3
Developing Erlotinib-Resistant Lung Cancer Cell Lines
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The human lung cancer HCC827 cell line was purchased from ATCC (ATCC® CRL-2868™). The PC9 cell line, which was derived from a human adenocarcinoma of lung tissue, was preserved in our laboratory. The lung cancer cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (Gibco BRL, Carlsbad, CA, USA) and 100 U/ml penicillin/streptomycin at 37°C in a humidified incubator containing 5% CO2. Erlotinib (OSI-744) was purchased from Selleck Chemicals (Houston, TX, USA). Insulin-like growth factor 1 human recombinant was obtained from ProSpec (ProSpec, Rehovot, Israel). Two erlotinib-resistant lines, namely HCC827/ER and PC9/ER, were developed by applying high-dose (1–5 μM) pulses of erlotinib combined with continuous low-dose (0.01 μM) administration for >8 months (13 (link)). To avoid the effects of the drugs, resistant cell lines were cultured in a drug-free medium for ≥2 weeks prior to further experiments.
4
NSCLC Cell Line Characterization
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NSCLC cell lines with EML4-ALK [27 (link)], H2228 (CRL-5935, ATCC, LCG standards, Wesel, Germany), and H3122 (Adi F. Gazdar, UT Southwestern, Dallas, TX, USA), or without EML4-ALK, HCC827 (CRL-2868, ATCC, LCG standards, Wesel, Germany), and A549 (CCL-185, ATCC, LCG standards, Wesel, Germany), were grown in an RPMI medium with 10% fetal calf serum and 1% penicillin–streptomycin solution. All cells were cultivated in 5% CO2 at 37 °C. Approximately 2.0 × 106 cells were lysed in 100 µL lysis buffer (Tris–HCl 50 mM, EDTA 10 mM, NP-40 1%, SDS 1%) and physically fragmented by sonication to an average fragment length of 200–500 bp. The DNA was treated with 20 µg Proteinase K and purified using the NucleoSpin Gen and PCR Clean-up kit (Macherey–Nagel, Dueren, Germany). The purified DNA was kept at − 20 °C until applied to NGS.
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