Testing the Effects: A Comparison of Culture Media for In Vitro T Cell Expansion and Function
Identifying a reliable culture media to support in vitro T cell studies is an important link in the chain of various immuno-oncology strategies.
We believe it is prudent to identify alternative media that can perform suitably in different conditions and for varied purposes. To address this issue, we have conducted a series of studies comparing the performance of several culture media.
Methods
We compared a variety of culture media (including classic media + supplements as well as new media) to several commercially available T cell media in the generation of primary MLR (mixed-lymphocyte reaction), antigen-recall assay (e.g., CMV, tetanus), antigen-specific T cell proliferation assay, and in anti-CD3/CD28-driven T cell expansion culture.
Results
We found that classical media supplemented with several defined components can support primary in vitro responses as measured by cytokine production. However, sustained T cell proliferation demanded additional supplementation and revealed greater differences between media.
This experiment demonstrates the effect of human AB serum (HS) or fetal bovine serum (FBS) added to the culture medium X-VIVO™ 15 (Lonza, Walkersville, MD). At low peptide concentrations (3 and 10 ng/mL), the presence of HS and FBS inhibits T cell proliferation compared to X-VIVO™ 15 alone, as seen in Figure 3.