We are working on HFF cell culture , but until now, the cell growth is very slow. The culture medium DMEM (1X)+GlutaMAX; + 4.5g/l D-glucose; - pyruvate ; 10%FBS and 1%antibiotic. Please how can we manage it? The incubation is at 37 degre Celsus and 5%CO2.
Hi Lauve,
- Check your culture medium if all supplements are added in accurate amounts.
- Also check the incubation conditions. Is the incubator at 5%CO2 atmosphere?
- What sub-type of HFF are you trying to grow? Go to the manufacturer website to check the culture conditions for your cell type.
Good luck
Fabrice
Thanks Patrick for your contribution. The culture is not infected.
Hi Lauve,
If the culture is not infected you should check the cells incubation atmosphere. Is it at 5%CO2 or so?
More than 10% Fetal Calf Serum (high purity, decomplemented at 56°C for 30 minutes and thawed less than 3 times) seems not to improve mitotic capacities of fibroblasts.
Human Foreskin Fibroblasts are from a primary culture that you started in house after dissecting human newborn tissues? If not, it could happen that your provider sent cells that were subjected to numerous passages before delivery. After each passage, the cell telomere sizes change and cell ability to grow is reduced.
My advice is to ask your cell provider the date of primary culture and the number of passages of the batch you are working with. It was shown that after 20 to 25 passages the mitotic index of several non-transformed cell lines is extremely low.
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