Tc20 automatic counter

Cells in a single-cell suspension were counted with a TC20 automatic counter (Bio-Rad, Hercules, CA, USA). Equal numbers of cells were cultured in triplicate on 24-well plates (CytoOne, USA Scientific, Ocala, FL, USA) for 7 days. Cell proliferation was assessed every 2 days by crystal violet staining, as we described previously [23 (link)].

As described, HT-29, DLD-1, HCT-116, Lovo, SW403 and SW48 human CRC cell lines were provided by Shanghai Institute of Biological Science (Shanghai, China). FHC colon epithelial cells were purchased from the iBS Fudan Cell Bank (Shanghai, China). Cells were maintained in DMEM plus 10% fetal bovine serum (FBS). The number of viable cells (trypan blue exclusive) was recorded via the TC20 automatic counter (Bio-Rad, Shanghai, China). All cell lines utilized in this study were subjected to mycoplasma and microbial contamination examination every two months. Population doubling time, colony forming efficiency, and morphology were also examined routinely for cell authentication.