Hi, everyone. I've been culturing hFOB 1.19 for the last two months. And at first after frozen the cells, they grow well and fast, but after the first subculture the cells starting to grow too slow. I had one flask of 25cm and it's been 2 weeks and they can't reach the confluence.
I grow the cells at 33°C with DEMEM:F12
Do you have any suggestions for supplements or something like that?
Hello Marian
The cells grow slowly when there seems to be contamination (especially mycoplasma contamination). Please check.
When cells are not frozen in a proper manner on thawing they may tend to grow slow. You need to do a viability check after thawing as it will give you an idea of the health status of the cells.
What is the passage number of the cells in culture? Cells with higher passages grow relatively slower than those which are in early passage.
Variation in culture condition like
the culture medium, serum lot, pH of medium can lead to slow growth. Please recheck all these parameters.
Have you added L-glutamine in the complete medium? The temperature for optimum growth for these cells is 33.5 degree celsius.
Do not subculture cells when they are over confluent.
I hope my suggestions will be helpful.
Good Luck.
Malcolm Nobre Thanks Malcolm, the passage number is 4, I will check about mycoplasma contamination and elevate the temperature to 33.5°.
I appreciate all the suggestions!
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