THP-1 was grown in T75 culture flasks (90075, TPP Switzerland) containing RPMI 1640 medium (R7388, Sigma-Aldrich) supplemented with 10% fetal calf serum (FCS, F9665, Sigma Aldrich), 1% l -glutamine (D7513, Sigma Aldrich) and 1% Penicillin Streptomycin Neomycin mix (P4083, Sigma Aldrich) at 37 °C in a humidified atmosphere of 5% CO2. For routine maintenance, THP-1 was subcultured and the medium was refreshed every 3–4 days. The Suspension cells were counted using an automated hemocytometer (Casy TT ® cell counter, Roche Innovatis AG). Subcultures were seeded at density 0.5–1 × 106 per 20 ml fresh medium at 37 °C in a humidified atmosphere of 5% CO2.
T75 culture flasks
Manufactured by Merck Group
Sourced in Switzerland, Germany
The T75 culture flask is a laboratory item used for cell culture and tissue culture applications. It provides a surface area of 75 cm² for the growth and maintenance of cells. The flask is made of tissue culture-treated polystyrene and features a vented cap to allow gas exchange.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Dulbecco s mem, by Harvard Bioscience (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
2 protocols using t75 culture flasks
1
Culturing and Maintenance of THP-1 Cells
2
Immortalized Human Corneal Endothelial Cells
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Cell cultivation SV40 immortalized human corneal endothelial cells from a 91-year-old Caucasian woman, initially generated by Bednarz et al. (2000) ; Nitschke et al. (2007); Valtink et al. (2008) , were acquired from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures (HCEC-12, Braunschweig, Germany). They were cultured in medium (M1), consisting of 95% DMEM (Dulbecco's MEM, Biochrom GmbH, Berlin, Germany) supplemented with 5% FBS (Fetal Bovine Serum, GE Healthcare, Freiburg, Germany), 1/100 Penicillin-Streptomycin (Sigma-Aldrich, Munich, Germany) and 1/125 Amphotericin B (Sigma-Aldrich), in T75 culture flasks (Sigma-Aldrich) at 37 °C in an incubator with 5% CO 2 (BBD 622, Thermo Scientific, Waltham, MA, USA). They were passaged every 7 days using 1 ml Trypsin-EDTA (0.05%; Gibco Invitrogen, Darmstadt, Germany) for 3 min. Trypsin activity was interrupted by adding 5 ml of M1. Cells were diluted in a 1:10 ratio.
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