After the samples were transferred to the 48-well tissue culture plates (Corning, UK), HCEC suspensions were seeded onto the β-CD-DA/OFLX-Col films (experimental group, n = 5) and the β-CD-DA-Col films (control group, n = 5). The seeded cell density was 1 × 104 cells per mL. After 1, 3, and 5 days, the proliferation activity of HCECs on the films was determined by a CCK-8 assay (Dojindo, Kumamoto, Japan) with a Thermo3001 microplate reader (Thermo, USA) using an optical density (OD) of 450 nm.
Thermo3001 microplate reader
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Thermo3001 microplate reader is a compact and versatile instrument designed for absorbance measurements in microplates. It can be used for a variety of applications, including enzyme-linked immunosorbent assays (ELISAs), cell-based assays, and colorimetric endpoint assays.
Automatically generated - may contain errors
3 protocols using thermo3001 microplate reader
1
HCEC Proliferation on β-CD-DA Films
2
Effect of Microgrooves on C2C12 Proliferation
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A CCK-8 (Dojindo
Laboratories, Japan)
was used to determine the effects of microgrooves with different widths
(W100D50, W50D25, and W25D50) and depths (W50D100, W50D50, and W50D25)
on the proliferation of C2C12 cells at different time intervals. In
brief, samples from both the experimental and control groups were
transferred to 24-well plates, seeded with 1 mL of a C2C12 cell suspension
(1 × 104 cells/well), and incubated at 37 °C
in a humidified incubator with 5% CO2. After culturing
for 1, 3, and 5 days, 300 μL of DMEM and 30 μL of CCK-8
solution were added and incubated with the cells at 37 °C for
1 h. Subsequently, the medium supernatant was extracted for absorbance
detection using a Thermo 3001 microplate reader (Thermo, USA) (n = 6) at 450 nm. In addition, the adhesion rate of cells
on the surface of the groove substrates with different widths was
also evaluated by CCK-8 assay after culture for 1 day.
Laboratories, Japan)
was used to determine the effects of microgrooves with different widths
(W100D50, W50D25, and W25D50) and depths (W50D100, W50D50, and W50D25)
on the proliferation of C2C12 cells at different time intervals. In
brief, samples from both the experimental and control groups were
transferred to 24-well plates, seeded with 1 mL of a C2C12 cell suspension
(1 × 104 cells/well), and incubated at 37 °C
in a humidified incubator with 5% CO2. After culturing
for 1, 3, and 5 days, 300 μL of DMEM and 30 μL of CCK-8
solution were added and incubated with the cells at 37 °C for
1 h. Subsequently, the medium supernatant was extracted for absorbance
detection using a Thermo 3001 microplate reader (Thermo, USA) (n = 6) at 450 nm. In addition, the adhesion rate of cells
on the surface of the groove substrates with different widths was
also evaluated by CCK-8 assay after culture for 1 day.
3
Galactose Cytotoxicity in ATDC5 and Chondrocytes
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The cytotoxicity of various concentrations of galactose toward ATDC5 cells and chondrocytes was characterized on days 1, 3, 5, and 7 using Cell Counting Kit-8 (CCK-8, Dojindo, Japan) assays according to the manufacturer’s instructions. The cells were seeded in 96-well plates (Corning, United States) at a density of 2000 cells/well, ATDC5 cells were maintained in a normal medium, and both types of cells were supplemented with galactose at a range of concentrations. At the indicated time point, the cells were incubated for 4 h in a CCK-8 working solution at 37°C and 5% CO2. After incubation, the absorbance of the solution was measured at 450 nm using a Thermo 3,001 microplate reader (Thermo Scientific, United States). The IC50 curves were generated by adding different concentrations of galactose to the medium, and the cells were cultured for 72 h, followed by cytotoxicity measurements using the CCK-8 assay.
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