I use fetal calf serum in the media used to culture B cells. Would any of the serum components be antigenic and affect my experiments?
Not sure I understand your question correctly but 10% fetal calf serum has been routinely used in B cell culture medium for >35 years without any issues. It is possible that some component may activate the BCR in a rare B cell clone but if you have relevant controls I don't think it will affect your readout. However, FCS does contain complement proteins that may affect the immune response and it is standard practice to heat inactivate it before use.
Hi, can you specify if you culture primary B-cells or stable cell line ?
I don't think that the serum are "antigenic" as it's probably not the BCR that are involved!!
However, serum components activate B cell proliferation. So you have to select your FCS batch to avoid this issue
I think the ab inside the serum is antigenic because it act as.idiotype and b cell will produce anti idiotype antibody
Not sure I understand your question correctly but 10% fetal calf serum has been routinely used in B cell culture medium for >35 years without any issues. It is possible that some component may activate the BCR in a rare B cell clone but if you have relevant controls I don't think it will affect your readout. However, FCS does contain complement proteins that may affect the immune response and it is standard practice to heat inactivate it before use.
If your experiments include signaling studies it is also common practice to serum starve your cells for a certain time before you perform your experiment and stimulate them.
I for example incubate the cells in medium containing only 1% FCS for 1 h prior to my experiment and this procedure is delivering good and reproducible results for signaling outputs such as calcium flux and tyrosine phosphorylation.
Thank you for your replies. I culture both primary cells and cell lines, and use heat inactivated serum. My experiments involve stimulating the cells and studying cell signaling. But if deprived of serum, the cells do not seem healthy. I do not necessarily mean to ask about the antigenicity via the BCR; any signaling or activation in general.
I am not sure if antibodies would be present in the FCS because it is taken from the fetus. Complements are taken care of by heat inactivating it.
Did you see this article? it might be relevant.
J Cell Biochem. 2008 Apr 1;103(5):1379-93.
Calcium oxalate crystals in fetal bovine serum: implications for cell culture, phagocytosis and biomineralization studies in vitro.
Pedraza CE1, Chien YC, McKee MD.
Author information
Abstract
Cell culture methods and models are key investigative tools for cell and molecular biology studies. Fetal bovine serum (FBS) is commonly used as an additive during cell culture since its constituents promote cell survival, proliferation and differentiation. Here we report that commercially available FBS from different major suppliers consistently contain precipitated, calcium oxalate crystals-either in the monohydrate (COM) or dihydrate (COD) form. Mineral structure and phase identification of the crystals were determined by X-ray diffraction, chemical composition by energy-dispersive X-ray microanalysis, and imaging and measurement of crystal growth steps by atomic force microscopy-all identified and confirmed crystallographic parameters for COM and COD. Proteins binding to the crystals were identified by immunoblotting, revealing the presence of osteopontin and fetuin-A (alpha(2)HS-glycoprotein)--known regulators of crystal growth found in serum. Macrophage cell cultures exposed to calcium oxalate crystals showed internalization of the crystals by phagocytosis in a process that induced disruption of cell-cell adhesion, release of reactive oxygen species and membrane damage, events that may be linked to the release of inflammatory cytokines by these cells into the culture media. In conclusion, calcium oxalate crystals found in commercially available FBS are toxic to cells, and their presence may confound results from in vitro studies where, amongst others, phagocytosis, biomineralization, renal cell and molecular biology, and drug and biomaterial testing are being examined.
2007 Wiley-Liss, Inc.
PMID:
17879965
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