Hello, why do I use serum-free culture medium when Jurkat cells are activated with antibodies?

Hi Zirong,

Thank you for your question. There are three reasons why we chose to use a serum-free medium for activating Jurkat T cells with anti-CD3 and anti-CD28 antibodies in our study.

First, bovine serum contains active components that could unintentionally activate the cells during culture. Previous studies have shown that fetal bovine serum (FBS) can induce a strong and prolonged response in the ERK MAP kinase pathway, which could interfere with our analysis of MAPK activation dynamics (Thrane et al., 2001; Ley et al., 2003).

Second, both our observations and previous research have demonstrated that T cell viability remains high in serum-free medium, and key characteristics such as the production of Interleukin-2 (IL-2) and immunological synapse formation are preserved during short-term culture (Uittenbogaart et al., 1983; Bi et al., 2001).

Third, it is challenging to obtain the same high-quality serum batch consistently throughout the entire study. Variability between different batches of serum could affect the T cell response to stimulation, leading to a lack of reproducibility in our experiments.

References:

1. Bi, K., Tanaka, Y., Coudronniere, N., Sugie, K., Hong, S., van Stipdonk, M.J., and Altman, A. (2001). Antigen-induced translocation of PKC-theta to membrane rafts is required for T cell activation. Nat Immunol 2, 556-563.
2. Ley, R., Balmanno, K., Hadfield, K., Weston, C., and Cook, S.J. (2003). Activation of the ERK1/2 signaling pathway promotes phosphorylation and proteasome-dependent degradation of the BH3-only protein, Bim. J Biol Chem 278, 18811-18816.
3. Thrane, E.V., Schwarze, P.E., Thoresen, G.H., Lag, M., and Refsnes, M. (2001). Persistent versus transient map kinase (ERK) activation in the proliferation of lung epithelial type 2 cells. Exp Lung Res 27, 387-400.
4. Uittenbogaart, C.H., Cantor, Y., and Fahey, J.L. (1983). Growth of human malignant lymphoid cell lines in serum-free medium. In Vitro 19, 67-72.

Thank you,

Xudong