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The CRL-1405 is a laboratory instrument designed for cell culture applications. It is a water-jacketed CO2 incubator that provides a controlled environment for the growth and maintenance of various cell lines. The CRL-1405 features temperature and CO2 regulation to support optimal cell culture conditions.

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2 protocols using crl 1405

1

Culturing Human Gastric and Guinea Pig Fibroblast Cells

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The human AGS (CRL-1739) gastric adenocarcinoma epithelial cell line[56 (link)] and guinea pig fibroblasts (CRL-1405)[57 (link)] were obtained from the American Type Culture Collection (ATCC, Rockville, Md.). The cells were routinely grown as a monolayer in complete RPMI-1640 medium (cRPMI; Sigma St. Louis, MI, United States), containing 10% heat inactivated Fetal Bovine Serum (FBS; CytoGen, Łódź, Poland), 1% penicillin/streptomycin (Gibco, Zug, Switzerland), at 37 °C in a humidified atmosphere containing 5% CO2. The cells were passaged every seven days with 0.25% trypsin/0.02% EDTA (HyClone, Thermo Fisher Scientific, Waltham, MA, United States) and the medium was changed every 3-4 d.
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2

Guinea Pig Gastric Epithelial Cell Isolation

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Primary guinea pig gastric epithelial cells were obtained as previously described [54 (link)]. Briefly, the stomach was isolated from the guinea pig, homogenized and treated with 0.25% trypsin (BIOMED-LUBLIN, Lublin, Poland) for 15 min at room temperature. The cell suspension at a density of 2 × 106 cells/mL was seeded onto a 6-well plate (Becton Dickinson, Franklin Lakes, NY, USA) and cultured for 24 h (5% CO2, 37 °C) to allow the cells to adhere. The cells were cultured in a mixture of DMEM and Ham’s F-12 media (ratio 1:1; Sigma-Aldrich, Saint Louis, Mi, USA), supplemented with 10% fetal calf serum (FCS), 1% (N-2-hydroxyethylpiperazine-N-2-ethane sulfonicacid) (HEPES), penicillin (100 U/mL), streptomycin (100 µg/mL), amphotericin B (0.025 µg/mL), l-glutamine (2 mM/mL)epidermal growth factor (Sigma-Aldrich, Saint Louis, MI, USA) 0.01 µg/mL and 0.005% dexamethasone. The guinea pig fibroblasts (CRL-1405) were obtained from the American Type Culture Collection (ATCC, Rockville, Manassas, VA, USA) The cells were routinely grown as a monolayer in complete RPMI-1640 medium cRPMI(Sigma St. Louis, MI, United States) at 37 °C in a humidified atmosphere containing 5% CO2.Every 2–3 days, the medium was changed and the cells were passaged at 80–90% confluence.
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