Cell lines were purchased from the Korea Cell Line Bank (KCLB, Seoul, Republic of Korea). RAW 264.7 and HaCaT cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Welgene, Republic of Korea) containing 10% fetal bovine serum (FBS, Gibco, Waltham, MA, USA) and 1% penicillin-streptomycin (Gibco, Waltham, MA, USA) in a humidified atmosphere of 5% CO2 at 37 °C. Cytotoxicity was determined using a water-soluble tetrazolium salt-1 (WST-1) assay (Dogenbio, Seoul, Republic of Korea).
Wst 1
Manufactured by DoGenBio
WST-1 is a colorimetric assay used for the determination of cell viability and proliferation. The assay is based on the cleavage of the tetrazolium salt WST-1 to a colored formazan dye by mitochondrial dehydrogenases in viable cells. The amount of formazan produced is directly proportional to the number of metabolically active cells in the culture.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Welgene (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Live dead viability cytotoxicity assay kit, by Thermo Fisher Scientific (1 mentions)
Microplate reader, by Tecan (1 mentions)
Fetal bovine serum (fbs), by Welgene (1 mentions)
4 protocols using wst 1
1
Cell Culture Cytotoxicity Assay
2
Evaluating DPSC Viability on CSP-CPC Scaffolds
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The DPSCs were grown in 96-well plates filled with CSP-CPC scaffolds at a density of 2 × 104 cells/well. Cytotoxicity was evaluated using a water-soluble tetrazolium salt assay kit (WST-1; Dogen-bio, Seoul, Republic of Korea). After 1, 3, and 7 days of incubation, the cells were washed with PBS and incubated for 1 h in a medium with 10% WST-1 reagent. Absorbance was measured at 450 nm using a microplate reader.
To evaluate the adhesion and viability of DPSCs seeded on CSP-CPC scaffolds, the Live/Dead Viability/Cytotoxicity Assay Kit (Invitrogen, Waltham, MA, USA) was used. Once the medium was removed and samples were washed once with PBS, a new medium with dye solution containing 2 mM ethidium homodimer−1 and 4 mM calcein-AM was added to each well, followed by a 30-min incubation period. The live and dead cells on the scaffolds were observed using a fluorescence microscope (Nikon, Tokyo, Japan).
To evaluate the adhesion and viability of DPSCs seeded on CSP-CPC scaffolds, the Live/Dead Viability/Cytotoxicity Assay Kit (Invitrogen, Waltham, MA, USA) was used. Once the medium was removed and samples were washed once with PBS, a new medium with dye solution containing 2 mM ethidium homodimer−1 and 4 mM calcein-AM was added to each well, followed by a 30-min incubation period. The live and dead cells on the scaffolds were observed using a fluorescence microscope (Nikon, Tokyo, Japan).
3
Cytotoxicity Evaluation of CS/PEG CPC Scaffolds
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Different concentration w/v ratios (0% and 5%) of CS/PEG CPC samples were sterilized in an autoclave at 120 °C for 1 h and placed in a 96-well plate. The DPSCs were incubated in a 6-well plate at a density of 3 × 104 cells/well. After reaching 70–80% confluence, the cells were sub-cultured and placed on the CS/PEG CPC in the wells. The cytotoxicity was evaluated using a water-soluble tetrazolium salt assay kit (WST-1; Dogenbio, Seoul, Korea). After incubation for one and seven days, the cells were washed twice with PBS and incubated in medium with 10% WST-1 reagent for 1 h. The absorbance was measured using a microplate reader at a wavelength of 450 nm. Three samples from each group were measured, and the data are displayed as the mean ± standard deviation.
4
Cytotoxicity Evaluation of Polymeric Compounds
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The mouse muscle cell line L929 (Korean Cell Line Bank, Seoul, Korea) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Welgene, Gyeongsan, Korea) supplemented with 10% (v/v) fetal bovine serum (FBS; Welgene, Gyeongsan, Korea) and 1% (v/v) penicillin-streptomycin antibiotic solution in a humidified 5% CO2 incubator. Cells were seeded in 96-well plates at 3 × 103 cells/well and incubated for 24 h. Subsequently, cells were treated with growth medium containing different concentrations of F127 and CS-mPEG (2%, 0.2% and 0.02%). After 72 h, the cytotoxicity of the L929 cells was evaluated using s water-soluble tetrazolium salt assay kit (WST-1; Dogenbio, Seoul, Korea). The WST assay was performed according to the product instructions. After 1 h of incubation, the solution was transferred into a 96-well plate and analyzed at 450 nm with a microplate reader (Tecan, Männedorf, Switzerland).
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