I want to know the optimal conditions for culturing fibroblast cells.
What kind of fibroblasts are they? Standard concentrations for the fibroblasts I culture (equine dermal) is 10%, but I increase to 20% on thawing frozen cells or if they look unhappy or stop growing.
Fibroblast Culture Medium:
Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 11960) supplemented with 10% Fetal Bovine Serum (FBS; Invitrogen,), 2mM L-glutamine (Invitrogen, cat. no. 25030), and 0.1mM (0.7µl/100ml final media volume) 2-mercaptoethanol (Sigma, cat.
no. M7522). Antibiotics can be added at final concentrations of 50 units/ml
penicillin and 50 g/ml streptomycin (Invitrogen, cat. no. 15070). Fibroblast Culture
Medium is filter sterilized, stored at 4°C, and used for up to 2 weeks.
Hope that helps. Good luck.
I worked with fibroblast from primary culture, and we cultured them at 10%FBS
I have used 10% as well. In DMEM supplemented with 1% Penn/Strep and 2mM L-glutamine as well.
If you are going to isolate them through outgrowth method then I would suggest using 20% until you see the cells appearing on the surface, but if you are just passaging them then 10% FBS is pretty enough. (obviously the more the FBS, the faster and better the cells will grow, but Fibroblasts are robust and well growing cells even with 5% FBS!)
i'm using SW872 cells, i use DMEM with 10% deactivated FBS for routine maintenance. However, when you carry out experiments you should use 1% FBS to avoid overgrowth & interference in your assays.
Grow at 37 degrees, 5% CO2 and 95% humidity.
Supplement ur media with glutamine, strep-pen and an anti-fungal agent.
DMEM with 10% FCS is standart, but to defreeze cells you have to use 20% for 1-2 days. Primary fibroblasts are better in 15% FCS for longer cultivation without senescence.
I usually begin with 15% FBS, and then gauge whether I need to increase to 20% or decrease to 10%. Usually anywhere between 10-20% FBS is usually a good rule of thumb when growing fibroblast cell cultures.
I will continue with 15% for cultivation, but 10% for "standart" experiments (if you don't study the effect of FCS concentration).
generally for normal growth 10% FBS is enough. to revival cells 10- 20% of FBS is required . some add 5% of FBS to maintaining the cells so growth of cells decreases
If you are going to isolate them through outgrowth method then I would suggest using 10% with minimal essential amino acids until you see the cells appearing on the surface, but if you are just passaging them then 5-7% FBS is enough. Some time more the FBS, compete with the cells during cell culture.
For good cell proliferation you have to use 15% FBS for primary fibroblast culture, so that to avoid of fast differentiation. Nevertheless, 10% FBS is usual standart conditions for experiments, so that normally we use this concentration for experiments, except for some special situations.
In our lab we are using 20% FBS for our 3T3 cell cultures. Needless to say, they love it and double their number everyday.
Generally for good cell proliferation you have to use 15% FBS for Primary fibroblast culture & 10% FBS for Cell line.
10 to 20% of FBS worked the best for my primary fibroblasts. However, beware of the differences in your results, depending on serum concentration. It is also recommended to synchronize your cells before the experiments.
10 to 20% of FBS for Primary fibroblasts cells.10% of FBS for fibroblast cell line is best.
Use modified mediums and you can decrease the FBS concentration to 2% for primary fibroblasts.
FBS concentration from 5% to 10 % have worked best for me for primary dermal fibroblasts
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