For the cell culture of this cell line, I use DMEM High Glucose culture medium along with recombinant human insulin and FBS, but the cell growth is extremely slow and a lot of time must be spent to increase its density and number. I used glutamine before, but it was ineffective. Do you know a solution to increase the speed of cell growth??
Hello Reza Nosrati,
I suspect the slow growth of your cell line is due to sub-optimal growth conditions. I'd recommend checking the below conditions:
1. The temperature (37C with 5% CO2).
2. pH to be around 7.4.
3. If the cell line is under the above 2 conditions, it could be due to over-crowdedness in the medium, which inhibit optimal cell growth.
I hope this helps, all the best,
Nicolas
It is worth checking for mycoplasma infection which can often slow cell growth and do not use too much culture medium as increased vlume of medium also dilutes the growth factors and slows growth
Nicolas ZuXuan Xu
Thank you for your explanation, dear Nicholas.
I have to say that yes, I follow all the conditions you mentioned about cell culture, but I still did not get a good result.
I would suggest that if the media is OK, there are no rests of soap in your bottles, pipettes, etc the reason most likely is mycoplasma contamination. You can check it by PCR, either buying primers or if somebody in a lab around uses them they tell you the sequences and the amplification conditions.
If this is the case the best is that you get new cells that you check to verify that they are not contaminated. For cells from a cell line like T47D is not worth to try to decontaminate them (in particularly since you would have to sub clone them and check for phenotype afterwards)
Best
Jose A.
- Badges
- Science topic
- Similar topics
- Social Psychology
- Attitude