Fetal Bovine Serum – See What 40-years of Consistency Looks Like

Why Serum Consistency Matters to Cell Culture Health

 

 

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 provides key quality parameters 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.

    
Product Size Regular Heat-inactivated Gamma-irradiated Heat-inactivated & Gamma-irradiated
Premium Select FBS

500 mL

S11550

S11550H

S11550G

S11550GH

100 mL

S11510

S11510H

 

50 mL

S11595

S11595H

S11595G

S11595GH

Premium Select FBS

 

 

Consistent & High Quality FBS

Plating efficiency of cells
Viability studies using R&D Systems 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

HUVEC expansion
Viability studies using R&D Systems FBS

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.

    
Product Size Regular Heat-inactivated Gamma-irradiated Heat-inactivated & Gamma-irradiated
Optima FBS

500 mL

S12450

S12450H

S12450G

S12450GH

100 mL

S12410

S12410H

 

50 mL

S12495

S12495H

S12495G

S12495GH

Optima FBS

 

View Data of Serum Consistency

Plating efficiency of cells
Viability studies using R&D Systems FBS

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

Two lots of Optima FBS showed consistent expansion rates for ATDC-5 cells.
Viability studies using R&D Systems FBS

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.

    
Product Size Regular Heat-inactivated
Premium FBS

500 mL

S11150

S11150H

100 mL

S11110

S11110H

50 mL

S11195

S11195H

Standard Grade Premium FBS

 

High Performance Standard Grade FBS

Plating efficiency of cells
Premium 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

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 PE-conjugated CD90 (R&D Systems, Catalog # FAB2067P) and APC-conjugated CD45 (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