How do you guys prepare 10% FBS Medium? - (Nov/27/2008 )

I am new to cell culture.

The people in my lab just add 50ml of FBS and 5 ml Pen Strep in 500ml DMEM. They say that the difference in cell growth is minimal. But i was taught to keep using the same lot of FBS throughout my whole experiment. But if the effect of different concentration of FBS on cell growth is minimal, what is the point of keep using the same lot of serum?

Thanks!

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If a bottle of DMEM/RPMI is 500ml and you need to make up a 10% FBS/FCS solution .....then you need to add 55mls of FBS/FCS to the 500ml of media. If you do not then you are adding a 10% error into your experiment straight away.

You need to batch test serum......this is extremely important as there is MASSIVE batch variation .........and MASSIVE differences in FBS/FCS origins.

NZ>Austalian>North American>European>South American>British

You get what you pay for:

Same rank order as above:

£250>£200>£130>£70>£60>£40 approximately


If you look at Vaibility and growth then there will be no differences between the above. HOWEVER these are insensitive methods for looking at the "health of the cells". If you look at other cell markers, then you will see massive variations:

Oxygen Consumption
Enzyme Induction
Receptor Expression
Cytochrome C contant
Electron transport chain complexes

Kindest regards


Rhombus

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Rhombus, that is a damn fine answer, especially as I am a Kiwi!

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Dear Bob1,

I am glad you like the answer. NZ serum has always been the best in the world......I do not know why. However experimentally it is vital for us to have good cells. For an example, we have HEK cells Sodium channel expression..........with NZ serum as the only varaible, we find 60% greater expression in cells grown in NZ serum compared to other sources of serum.......IMPORTANT when you are doing patch clamp work

Kindest regards

Rhombus

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