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The cells were maintained in humidified 5% CO2 incubator and passaged according to the recommended dilutions and confluency by ATCC.
All mosquitoes, including larval and adult mosquitoes, were maintained in humidified incubators at 25 ± 1 °C on a 12-h light: dark photocycle.
Vero cells were cultured in DMEM/F12 supplemented with 10% FBS, detached through application of 0.25% trypsin/EDTA and maintained in humidified atmosphere of 5% CO2 at 37°C.
All differentiating NPC cultures were maintained in humidified incubators at 37°C (5% CO2 in air).
Flies were maintained in humidified, temperature controlled chambers at 25°C and 60% relative humidity and under a 12∶12 light∶dark cycle.
Cells were maintained in humidified air with 5% CO2.
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The cells were grown initially in 25-cm2 flasks containing Dulbecco's modified Eagle's medium (DMEM) with 10% fetal calf serum, 3 mM L-glutamine, 1 mM nonessential amino acids as well as 50 units/ml penicillin/streptomycin (DMEM and cell culture additives were purchased from Invitrogen, Carlsbad, CA), and were maintained in a 5% humidified incubator at 37°C.
The cell cultures were maintained in a water humidified 37 °C incubator with 5% CO2.
The CM DGs were maintained in DMEM/F12 containing 10% FBS at 37°C in a 5% humidified CO2 atmosphere.
The human cholangiocarcinoma cell lines, TFK-1, HuCC-T1 and MEC were provided by the Cell Resource Center of Biomedical Research, Institute of Development, Aging and Cancer Tohoku Universityy, Sendai, Japan) and maintained in RPMI 1640 medium containing 10% fetal bovine serum (FBS) and antibiotics at 37°C in a 5% humidified CO2 atmosphere.
T47D cells were maintained in RPMI-1640 media containing insulin (5 μg/ml), FBS (10%), and penicillin/streptomycin (1%) at 37°C in a 5% humidified incubator.
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