SHZ-88 mammary gland carcinoma cells were purchased commercially from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Sangon Biotech, Shanghai, China) in a 37°C constant-temperature incubator (Thermo, USA) containing 5% CO2. The cell culture medium was changed every two days, and cell passage was conducted every week. In order to detach, the cultured cells were digested by 0.25% trypsin for 5–8 min, and the enzyme reaction was terminated with a culture medium containing 10% fetal bovine serum. For cells harvesting, the cells and medium were centrifuged for 3 min at 1000 r/min. The supernatant thus obtained was discarded and the pellet resuspended in the RPMI-1640 medium and diluted to achieve a final concentration of 107 cells/0.5 mL. The cells were counted by using a hemocytometer. Cell viability was estimated by MTT ((3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)) assay as described previously.37
37 c constant temperature incubator
Manufactured by Thermo Fisher Scientific
Sourced in China, United States
The 37°C constant-temperature incubator is a laboratory equipment designed to maintain a stable temperature of 37°C. It is used to provide a controlled environment for various applications that require a consistent temperature.
Automatically generated - may contain errors
2 protocols using 37 c constant temperature incubator
1
Culturing SHZ-88 Mammary Carcinoma Cells
2
Measuring Cell Proliferation with CCK-8
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Cell Counting Kit-8 (CCK-8, Beyotime, Shanghai, China) was used to detect cell proliferation ability. Cells were seeded in 96-well plates (5000 cells/well) and cultured for 24, 48, 72 and 96 h. For the dosing group, an additional 6.25 μmol/L PD98059 was added to each well. The medium in the wells was then discarded. Reconstituted fresh medium containing 10% CCK-8 was added to the 96-well plate in an amount of 100 μL per well. The 96-well plate was placed in a 37 °C constant temperature incubator (Thermo Scientific, Waltham, MA, USA) for 1 h. Finally, the absorbance value of each well at 450 nm was measured with a microplate reader (Thermo Scientific, Waltham, MA, USA).
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