Technical problems working with HCT116 compared with other cell line Caco-2 and HT29
Hello Y. Sangeeta
The technical issue with HCT116 cells is that these cells do not form a monolayer but grow in layers if allowed to grow unchecked in the flask. I feel that these cells have no contact inhibition. So do not allow the cells to become over confluent. If allowed to overgrow they will form clumps which will unnecessary add extra steps at the time of subculturing. You will have to vigorously pipette up and down to break the clumps into single cells after trypsinization.
Use 0.25%(w/v) Trypsin- 0.53mM EDTA solution for trypsinization.
Avoid cells of late passage for your studies.
Ganapathy has already mentioned about the culture medium in which HCT116 cells have to be maintained.
I hope this information is helpful.
All the best.
Dear Y Sangeeta!
Please you also see in this article:
Article Maintenance of HCT116 colon cancer cell line conforms to a s...
Cell culture. The colon cancer cell lines WiDr, HT29, DLD-1, and HCT116 were cultured in DMEM ⁄ F12 media supplemented with 10% FCS, 100 units⁄mL penicillin G, and 100 lg ⁄mL streptomycin (Nacalai tesque, Kyoto, Japan). For floating culture, HCT116 cells were cultured in serum-free DMEM ⁄ F12 media containing 5 lg ⁄mL of bovine insulin, 0.4% BSA, 10 ng ⁄mL of b-FGF, and 20 ng ⁄mL of EGF in 100 mm ultra-low attachment dishes (Corning Japan, Tokyo, Japan) at a density of 2.0 · 104 cells⁄mL at 37C in a humidified 5% CO2 ⁄ 95% air atmosphere
https://jcs.biologists.org/content/joces/130/1/203.full.pdf (attached)
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