Recently I have been problem with HEK293a expansion problem, after 2 passages in flask, the cells stop growing and detach from flask. I am cultivating using DMEM 10 % FBS (High glucose, 10 mM HEPES, 0,2% sodium bicarbonate, 59 mg/L penicillin and 133 mg/L streptomycin) all from Sigma. I need to expand this cells to 16 flask of T-150 (Corning). I don't know what is wrong. I would love to hear with someone had a similar problem.
Hey Isau. I principle, HEK293 cells are very robust, but could be you split them too hard? How confluent are they after the problematic split? They need some contact with each other to grow in an optimal manner.
hey Inbal Ipenberg thanks for replying. I am splitting them by factor of 2, one flask of T-75 for 2 flask of T-75 or 1 flask of T-150. I usually check if the cell are enough for cell contact. The problem start around 90% confluence and this happen 2 times. is it possible my HEKs going through cell senescence?
Hey,
Are you using cells with high passage? Do you check for micoplasma regularly?
- Badges
- Science method