2.6 | Cell viability assays
LF Cells were seeded in 96-well plates at 105 cells
per well and cultured at 37℃ under 5% CO2 for 24 h.
Then, LF cells were incubated in 80 µl of DMEM adding 20 μl of 5 mg/ml
3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT)
solution (Beyotime, Nanjing, China) for 4 h. After the media removal,
formazan crystals were dissolved in 100 μl DMSO/well, and the absorbance
of the solution was measured at 490 nm. For colony-forming unit
detection, the cells were pre-treated with Crystal Violet Staining
Solution (Beyotime, Nanjing, China) for 10 min at 37˚C. For EdU and BrDu
detection, the cells were pre-treated with EdU and BrDu working solution
(Beyotime, Nanjing, China) for 2h at 37˚C. Finally, they were observed
under the microscope (OLYMPUS, Nanjing, China).