Serum is added as a supplement to culture media at a concentration of 2-10% to provide a mixture of nutritional, hormonal, growth and attachment factors. Serum also acts as a buffer to the cell culture system against a variety of disruptions to cell growth and toxic effects such as pH change, proteoloytic activity, or the presence of heavy metals or endotoxin.
Serum delivered frozen to a processing facility is thawed and tested for the presence of bacteria, virus, endotoxin and for hemoglobin content. The best quality serum is then filtered through a sequence of membrane filters, terminating with a 0.1 micron pore filtration under aseptic conditions. After filtration, aseptic conditions are maintained as serum is dispensed into sterile bottles, quickly frozen to -20 °C and quarantined until all lots have met or exceeded quality control standards. A certificate of analysis for each lot is available upon request.
The recommended storage temperature for serum is between -5 °C and -20 °C, as it is most stable when stored frozen until needed. Storing serum in a frost-free freezer is not recommended, because temperature cycling may cause bottles to crack, contributing to contamination or deterioration of the product.
Serum is thawed overnight at 2-8 °C to prevent degradation; or, it may be thawed at room temperature with periodic agitation to resuspend the contents. Thawed serum should be mixed thoroughly before being added to cell culture medium.
As repeated freeze-thaw cycles may significantly impact serum quality, we recommend aliquoting the thawed serum for single use, and to store aliquots at -20 °C. If serum is stored between 2-8 °C, it should be used within 2-4 weeks.
Note: Temperatures above 37 °C may degrade nutrient components and compromise serum performance.
Serum is packaged in dry ice and it should arrive frozen. However, if serum is partially thawed due to delay, it can still be used. Thaw the serum completely and aliquot into single-use aliquots for storage and further use.
The shelf life of serum is five years from the date of manufacture.
Normal handling of serum under typical room/laboratory lighting will have little effect. However, serum does contain certain constituents that are light sensitive. Exposing serum to high-intensity or short-wavelength lights is therefore not recommended.
Precipitates appear in serum for a variety of reasons:
If aseptic handling is carefully observed, serum-supplemented media may be stored up to six weeks at 2-8 °C. Media should be discarded using appropriate institutional biosafety protocols if it becomes cloudy, regardless of storage duration.
Like all of our serum products, FBS is meticulously tested for the presence of these relevant microbial contaminants. A Certificate of Analysis (CoA, CofA) for each lot is available upon request.
Yes. All Sigma-Aldrich sera have been sourced from countries that are free of or at low risk for BSE. The intergovernmental World Organisation for Animal Health (OIE) fights animal disease at the global level by monitoring country compliance with OIE guidelines and standards.
Gamma-irradiated serum has been exposed to a Cobalt-60 source at a 25-35 kGy dose to inactive viruses and other adventitious microbial agents (such as mycoplasmas). Gamma irradiation does not compromise the physiochemical properties or the cell culture performance of serum.
Complement proteins naturally occurring in mammalian sera mediate cytolytic events, contract smooth muscles, and activate lymphocytic and myeloid cells. Heat-inactivation destroys the activity of complement in serum. This method is therefore recommended for immunological applications and for the culture of embryonic stem cells (ESCs), insect cells and smooth muscle cells.
The heat inactivation process must be carefully controlled to prevent denaturation of desirable serum proteins that are key constituents of sera necessary for cell and tissue propagation.
Heat inactivation occurs when serum is heated to 56 °C for a period of 30 minutes.
As serum is a solution extracted from biological sources, its composition may vary from lot to lot. To minimize application variability, we perform rigorous quality testing of each lot, and can provide lot samples to our customers for testing in their labs. We reserve material from the test batch during the customers’ lot qualification process.
The color of the serum depends on hemoglobin concentration, and variation in this parameter will not affect serum performance.
The origin of serum (also sometimes referred to as the ‘source’) is the country where the unrefined blood was collected. This is not to be confused with the country where the serum was separated, filtered, or otherwise processed, which may differ. The certificate of origin documents from the serum supplier should clearly state the serum source and processing information.