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CO2 incubator

Manufactured by Astec
Sourced in Japan

The CO2 incubator is a laboratory equipment designed to provide a controlled environment for cell and tissue culture applications. It maintains a consistent temperature, humidity, and carbon dioxide (CO2) concentration to support the optimal growth and maintenance of cultured cells.

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5 protocols using CO2 incubator

The skin fibroblast used in the experiments was NHDF-c (adult human dermal fibroblast) obtained from Promo-Cell (Germany). Skin fibroblast was cultured in DMEM media (Welgene Inc., Korea) supplemented with 10% fetal bovine serum (Gibco BRL, USA) and 1% penicillin/streptomycin (Welgene Inc., Korea) at 37℃ in a 5% CO2 incubator (ASTEC, Japan). The used cell passage numbers is between three and ten. Skin fibroblast was cultured for 24 h until they reached 70% confluency, and then further cultured in the FBS-free media for 24 h. Each infusion of Hanwoo’s leg bone, foot and tail was added at the DMEM media without FBS, cells were cultured in 5% CO2 incubator (ASTEC, Japan) for 48 h. The control group was cultured in the media without the infusions.
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2

Evaluating Biocompatibility of Drug-Loaded Nanoparticles

The biocompatibility of the drug-loaded triple-ion-substituted nanoparticulate system was tested against Swiss 3T3 fibroblast cells (NCCS, Pune) by the colorimetric MTT [3-(4, 5-181dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide] assay for 48 h. The Swiss 3T3 fibroblast cells were grown to confluence with Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% 100 × antibiotic–antimycotic liquid and incubated at 37°C with 5% carbon dioxide in a CO2 incubator (Astec, Japan). The cells were then trypsinized and counted with a hemocytometer (Marienfeld, Germany). They were then diluted at 5,000 cells per well, seeded in 96-well plates, and cultured for 24 h. Ten milligrams of the samples was suspended in 1 ml of DMEM and incubated at 37°C for 24 h. The media in the 96-well plates was then replaced with 100 μl of the supernatant from the CDHA samples and again incubated for 24 h. Twenty microliters of 5 mg/ml MTT was added to each well and incubated for 4 h. The purple formazan precipitate formed selectively by live cells was solubilized in dimethyl sulfoxide (DMSO), and the absorbance was measured at 570 nm using a multimode plate reader (EnSpire, Perkin-Elmer, Singapore). The percentage of viable cells was calculated as the percentage relative to the control (standard polystyrene tissue culture plates) using the following equation:
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Primary hAD-MSCs were used to investigate the influence of HPGs as scaffolding building blocks on the behavior of the cell. HPGs (2 mg) were placed in each well of a 24-well plate and sterilized using 70% (v/v) ethanol solution for 1 h, then dried and washed twice in PBS for 30 minutes. The cells were seeded at a density of 3500 cells per well and incubated with the basal medium of DMEM/F12 containing l-glutamine/10% FBS/1% antibiotics at 37°C in a humidified atmosphere of 5% CO2 incubator (ASTEC, Fukuoka, Japan). The medium was replaced every 48 hours.
To investigate the synergistic differentiation behavior, two test groups of HPGs and HPGs-BMP4 were used. Osteogenic differentiation culture medium was constituted of the basal medium supplemented with dexamethasone (10 nM), β-glycerophosphate (10 mM), and ascorbic acid (50 μM).
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L6 myoblast cells were cultured in DMEM (Medium contains DMEM with 10% FBS and 1% of 100 X antibiotic-antimycotic liquid) in a CO2 incubator (Astec, Japan) (temperature 37 °C, carbon dioxide level 5 %). Approximately104 cells were seeded into a 96 well polystyrene plate and incubated for 24 h. The cells were then incubated with 500 μg/ml of CβG/PLGA nanoparticles for another 24 h. Total number of viable cells was determined by MTT (3-(4, 5- Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) based cell proliferation assay [25] (link).
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The cell proliferation rate was measured by using the Cell Proliferation Reagent WST-1 (Roche, Germany). The skin fibroblast was cultured in 96-well plates (4×103 /well) in DMEM media with10% FBS for 24 h and cultured additionally in the media without FBS for 24 h. Next, each infusion was added to the DMEM media without FBS and skin fibroblast was cultured in 5% CO2 incubator for 72 h. Skin fibroblast was cultured at 37℃ in a 5% CO2 incubator (ASTEC, Japan) for 4 h after adding 10 uL of Cell Proliferation Reagent WST-1 to each well. Absorbance was measured at 450 nm by using of ELISA Reader (Molecular Devices, UK).
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