Why Serum Consistency Matters to Cell Culture Health
A case study using R&D Systems Fetal Bovine Serum
Introduction
For many cell lines, primary, and tissue culture systems fetal bovine serum (FBS) is relied upon for optimizing cell health and performance. The nutrients provided by FBS have a critical impact on cell culture reproducibility, including effects on cell expansion, viability, attachment, and cell-specific performance assays. Under these circumstances, the quality and consistency of FBS needs to be top of mind for the researcher.
This application note discusses key quality parameters to consider when sourcing your FBS, and provide data illustrating the 40-year historical consistency of our FBS products.
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Key Quality Tests for FBS Consistency
Plating Efficiency
Plating efficiency is used to assess the ability of serum to facilitate cell adherence to solid surfaces for growth. One common assay for this uses A549 human lung carcinoma cells, plated at low density in media supplemented with 5-10% FBS, and monitored for attachment and clonal growth after 10-14 days. This test is performed on all grades of R&D Systems FBS.
Cell Expansion and Viability
This standard test is used to evaluate the nutritional support of the FBS and is typically evaluated on both substrate-bound (adherent) cells and cells grown in suspension. High grade FBS should report high and consistent expansion and viability across lots and across cell lines. Cell expansion and viability tests are performed on all grades of R&D Systems FBS.
Hybridoma Cloning
The hybridoma cloning test measures the ability of a serum lot to support clonal growth of a hybridoma or myeloma cell line. Typically, a hybridoma cell line is plated at low density in a 96-well plate containing media supplemented with 5-10% FBS. High grade FBS should see high and consistent levels of cloning efficiency. R&D Systems FBS keep monitored for hybridoma cloning efficiency.
Cell-specific Assays
Researchers often need to self-evaluate FBS for performance in their specialized cultures. This process is simplified when the serum being tested is already quality-vetted by the manufacturer for consistency in plating efficiency, cloning, expansion, and viability.
Expected FBS Quality Specifications
For each lot of FBS, the following specifications should also be expected for FBS.
Endotoxin
Minimizing endotoxin levels in FBS improves performance. It is typically evaluated using LAL gel-clotting assay.
Mycoplasma
Triple 0.1 µm filtration removes mycoplasma from serum. Tests verifying the absence of mycoplasma should be reported for FBS.
Biochemical Profile
Protein and chemical profiling helps track lot consistency (testing regiment may vary by FBS type).
Hormone Profile
Performed on high-grade serumto elevate product consistency and transparency for researcherslooking to manage hormone influence in their cultures.
Viral Testing
Ensures serum is free ofspecific viral agents (testing regimentmay vary by FBS type).
Hemoglobin
Ranges from 10 – 30 mg/dL in FBS and is typically measureby spectrophotometry.
Premium Select FBS
Premium Select FBS undergoes extensive hormone profiling and rigorous biochemical analysis and viral testing, making this our highest-grade serum. This FBS is ideal for all cell culture applications, including culturing demanding or sensitive cell lines where consistent product profiling and lot-to-lot performance is critical.
Examples of performance consistency, both in-house and in the field, are shown below.
Consistent & High Quality FBS
Premium Select FBS Performs Consistently Across Lots and Performance Assays. Quality control data for Premium Select FBS shows high levels of performance consistency. (A) Plating efficiency of cells cultured in media containing 5% or 10% Premium FBS, relative to control lots of FBS. (B) Premium Select FBS is formulated to consistently support hybridoma cloning and is qualified for consistent and healthy expansion of adherent and suspension cell culture lines. In addition to its extended hormonal and biochemical profiling, lots of Premium Select FBS show consistent and high percentage cloning efficiency and cell viability.
Performance in the Field
Expansion of Human Umbilical Cord Endothelial Cells (HUVECs) using Premium Select FBS. HUVECs were expanded for 11 passages in media containing Premium Select FBS or an alternate supplier of FBS. (A) HUVEC expansion was consistent over 11 passages and comparable to Supplier #1. (B) Representative images of HUVECs after 9 passages in culture.
Optima FBS
The lot-to-lot consistency of Optima FBS, along with its extended biochemical analysis and APHIS-certified manufacturing process, makes this serum ideal for both standard and regulated cell culture applications. Similar to our Premium Select FBS, Optima undergoes extensive quality control testing to ensure consistent product specifications and performance. Each lot of Optima FBS is manufactured exclusively from US-origin material following stringent manufacturing and quality control procedures that fall under a USDA-APHIS certified process.
Examples of performance consistency both in-house and in the field are shown below.
View Data of Serum Consistency
Optima FBS has Consistent Lot-to-Lot Performance. Quality control data is displayed across multiple lots of Optima FBS. (A) Plating efficiency of cells cultured in media containing 5% or 10% Optima FBS, relative to control lots of FBS. (B) Optima FBS was tested for its ability to support hybridoma cloning as well as the healthy expansion of adherent and suspension cell culture lines. Lots of Optima FBS show consistent and high percentage cloning efficiency and cell viability.
How Does Optima FBS Perform? View Our Healthy Cell Data
Optima FBS Support Expansion and Health Across Cell Lines. Ba/F3 murine pro-B cell line, ATDC-5 murine teratocarcinoma cell line, and a TF-1 erythroleukemic cell line were evaluated for expansion and health in culture media supplemented with either Optima FBS or FBS from an alternate supplier (Suppler #1). (A) Two lots of Optima FBS showed consistent expansion rates for ATDC-5 cells. The expansion was similar to FBS from another supplier. (B) Ba/F3, ATDC-5, and TF-1 cell lines cultured in Optima FBS show consistent cell viability across multiple lots. The viability was comparable to an alternate supplier of FBS.
Standard Grade Premium FBS
Premium FBS is the most widely used grade of serum because of its combination of affordability, high performance, and consistency. Premium FBS undergoes more basic biochemical profiling but maintains of the same level of high-performance consistency as our other FBS grades, making this serum ideal for maximizing value.
Examples of performance consistency both in-house and in the field are shown below.
High Performance Standard Grade FBS
Premium FBS Displays Lot-to-Lot Consistency. Quality control data for over 80 lots of Premium FBS shows high levels of performance consistency. (A) Plating efficiency of cells cultured in media containing 5% or 10% Premium FBS, relative to control lots of FBS. (B) Premium FBS was tested for its ability to support hybridoma cloning as well as the healthy expansion of adherent and suspension cell culture lines. Lots of Premium FBS show consistent and high percentage cloning efficiency and cell viability.
Performance in the Field
Growth of MSCs using Premium FBS. Human mesenchymal stem cells were cultured for 4 passages in media supplemented with Premium FBS or an alternate FBS supplier (Supplier #1). MSCs were analyzed by flow cytometry using a PE-conjugated Mouse Anti-Human CD90 Monoclonal Antibody (R&D Systems, Catalog # FAB2067P) and an APC-conjugated Mouse Anti-Human CD45 Monoclonal Antibody (R&D Systems, Catalog # FAB1430A). (A) Percentage of CD90+ MSCs following four passages in culture. (B) Representative flow cytometry data of CD90+ human MSCs cultured for 4 passages in Premium Grade FBS.
Fetal Bovine Serum Selection Guide
Premium Select
Serum for demanding and sensitive cell lines
- Extensive hormone, biochemical, and viral testing profiles
- Manufactured from US- or USDA-approved origin
- Triple 0.1 μm filtered
Optima
For standard and specialized cell culture applications
- Extended biochemical profiling and viral testing
- Manufactured using US origin material under a USDA-APHIS certified process
- Triple 0.1 μm filtered
Premium
For general cell culture
- Basic biochemical profiling and viral testing
- Maximizes value and performance
- Manufactured from USDA-approved origin material
- Triple 0.1 μm filtered