Alternative Sera for Cell Culture - (Jul/24/2007 )
The lab that I have been working in for the last 3 years uses only Fetal Bovine Serum for cell culture media supplementation. There are several other variety of sera available, at significantly lower cost. Some prices I found were approximately:
Fetal Bovine Serum: $250 for 500mL
Fetal Calf Serum: $250 for 500mL
Bovine Serum: $40 for 500mL
Horse Serum: $40 for 500mL
I searched around on the internet a bit, but couldn't find a clear reason that the sera of young animals is enough superior for cell culture than that of their seniors. I don't doubt that it is, as I seldom see papers in my line of work listing the use of anything but FBS or FCS. I could see why it is more expensive to acquire, being less of it, but why is it better to use thus justifying the cost?
I found some answers to my own post by finding some up to date, inquisitive web postings (one excellent link below), and by calling ATCC and asking about the differences in the serum, and asked about heat inactivation of serum. It turns out that the most important thing I probably learned today is that I should not be heat inactivating my FBS. The advantage of FBS is more abundant growth factors, which are destroyed by the heat inactivation I have been applying to it. So heat inactivating the expensive FBS yields something kin to the cheap serum. ATCC said that some cell types do require heat inactivated FBS, but most do not.
The post below is very nice, and I think that many of you may also be unnecessarily either heat inactivating FBS that would be better without doing that, or could buy the cheaper serum. Much appreciation to someone at the UNC Tissue Culture Facility for there excellent post, and to ATCC technical support.
Matt
http://www.unc.edu/depts/tcf/tech_tipsHI.htm
Thanks a lot! It was a very useful link!
The post below is very nice, and I think that many of you may also be unnecessarily either heat inactivating FBS that would be better without doing that, or could buy the cheaper serum. Much appreciation to someone at the UNC Tissue Culture Facility for there excellent post, and to ATCC technical support.
Matt
http://www.unc.edu/depts/tcf/tech_tipsHI.htm
Things to know about FCS/FBS :
They are both the same thing
Because of Ultrafiltration HEAT INACTIVATION is no longer required to destroy compliment and if done will only denauture vital growth factors for cell growth.
New Zealand based FCS/FBS is the best serum in the world and therefore the most expensive.
It can be stored for many years without losing it's ability to support growth. So if a company comes to you with an offer on serum which is coming to it's sell by date....BUY IT.
It should be stored at -80oC. When thawing it is recommended to put it from -80oC into a fridge (+4oC) to slowly thaw. It is recommended to aliquot the 500ml of thawed FCS/FBS into usable amounts i.e. 55ml. It can then be re-frozen and stored short term at -20oC.
ALWAYS ALWAYS BATCH TEST and then buy a large amount to cover your research for at least 2-3 years.
DO A SENSITIVE TEST FOR BATCH TESTING....not viability or growth curves which are very insensitive.
Hope this is useful
Rhombus
I know this maybe variable for different purposes, what exact did you mean SENSITIVE TEST? Could you give us an example?
Dear Genehunter-1,
Sensitive tests in our lab include:
i) Oxygen consumption of the cells (Jurkats) i.e. when the cells are dying, in sub-standard FCS/FBS or drug insult, oxygen consumption falls rapidly ( as measured by Clark Electrode ).
ii) cGMP concentration within smooth muscle cell lines i.e. > in cGMP concentration is important in the modulation of vascular tone and causes > in intracellular Calcium release.
iii) Sodium Channel expression in transfected cells (HEK cells) i.e. if the cells are in sub standard serum then expression levels will drop.
iv) Inducibilty of Nitric Oxide synthase i.e. I can induce iNOS in J774.A1 cells by stimulation with LPS and Interferon gamma. In sub-standard serum I get 70% less expression when compared to New Zealand serum.
All the above tests are used in our batch testing. We want the same results from one year to the next and therefore reduce variability. If you just do growth and viability you will see NO difference between batches of serum or countries of origin.
New Zealand serum ALWAYS out performs all other serums. The quality is usually in this order:
NZ>Australian>US>South American>European>British
Price is also in the same order:
£250/£200/£150/£100/£50/£30.
We found recently that some cell lines can be grown in synthetic serum-- so in place of using FBS we switched to Fetalplex. It was very economic and did not require heat inactivation.
But some cell lines did not respond well to it, e.g. SHSY-5Y