Hello,
I am working with Mouse Anti-TNP-KLH Hybridoma [2C8.22] (CSCH0878) cell line for my project. I thawed cells a week ago but they are growing very slowly and the cell medium color is still pink. The medium I used is RPMI 1640 with 10% FBS and I am planning to change the medium and add RPMI+8%FBS, then I will gradually decrease the concentration of FBS in the medium as I need the cells to measure the antibody production.
My question: is there any recommendation to let the cells grow faster and which medium you might recommend for better growing of my cell line?
Thank you!
I don't think there is anything wrong with your choice of medium or FBS concentration. Some people use DPMI. Stick with 10% FBS until cells are growing quickly. I used to supplement with glutamine and pyruvate as well, but it is not unusual to have slow growth initially. I suspect you have a recovery issue from liquid nitrogen and thus changing to a different medium won't actually help.
Have you checked cell viability? It's possible that you have low viability and hence the medium is not being conditioned quickly. If so, I would let the cells settle and remove half of the medium to increase the concentration of live cells per ml. Once the medium starts to turn more orange/yellow, add more medium but do it more gradually than you would with a really healthy culture.
Finally, it is worth checking that your CO2 incubator calibration is correct. If this is awkward with cells in the incubator, you can simply place a spare flask of medium in the incubator overnight, and then check the pH with a standard pH meter the next day.
Actually the cell culture medium I used is RPMI+10% FBS, 2 mM L-Glutamine, 1 mM sodium pyruvate, 4500 mg/L glucose and for sure antibiotics and anti fungal. I will wait and see how they will grow. Thank you again!