I would like to find out if anyone has attempted to maintain S2 cell cultures using Schneider's medium supplemented with calf serum or newborn calf serum, which I've found are cheaper alternatives to FBS.
Thanks!
Hi, new born calf serum works fine for these cells. I have used that previously
Good Luck
Thanks! Do you use calf serum during transfection experiments as well?
By the way, I have found that heat inactivation of FBS is unnecessary for growing S2 cells. Do you know if this this also true for newborn calf serum?
The primary objective of heat inactivation is to inactivate heat labile components of the serum, primarily complement system. In addition, heat inactivation was introduced to neutralize microbes that can interfere with culture systems. Whether to heat inactivate a serum or not still remains a debatable issue. For S2 cells, I used heat inactivated serum just because we purchased it commercially and were using it even for mammalian cells. However, I do not foresee any problem even if you do not heat inactivate your serum. I am sorry I could not give you any definitive answer. Also, I never used serum when doing transfection. However, 6-12h following transfection, I changed media to one that contained serum.
http://depts.washington.edu/taneli/documents/protocols/schneidercells_man-1.pdf
Here is a link to an article about culturing and transfection of S2 cells. I believe you already have it.
Yup, thanks!
That's what I wanted to know concerning your transfection experiment - whether you used calf serum or FBS during the recovery phase.
I've read that the inactivation temperature is too low for neutralizing mycoplasmas, and complement proteins don't seem to negatively affect most cells. I've so far found that my S2 cells grow slightly faster and achieve confluency more rapidly in medium containing non-inactivated serum. Hence, I am going to dispense with heat inactivation and the added cost for FBS.
http://fscimage.fishersci.com/cmsassets/downloads/segment/Scientific/pdf/Cell_Culture/Application_Notes/heat_inactivation.pdf?&storeId=10652
I have observed this phenomenon with quite a few cell lines. For some reason unknown to me, heat inactivated serum seems to slow down growth. One reason might be that heating serum destroys a few critical components in the serum that we are unaware of. Anyways, good luck.
In my previous lab, we used 10% heat-inactivated FBS in our culture medium for Drosophila S2 cell passaging as well as transfections.
Hi Breanna,
My testing agrees with V. Anaparti's observation that non-inactivated FBS promotes faster proliferation of the cells.
Growth in medium supplemented with 10% newborn calf serum is also slower than in medium supplemented with 10% FBS but this may be advantageous for routine maintenance.
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