Hii everyone,
I am interested in treating primary cells with some cytokines. I found these cells do not grow properly in serum free media. However, literature suggests the treatment should be given in serum free media.
Please advice.
Hi, let the cells grow in serum-containing medium until they reach the needed confluency. Change medium to serum-free medium, wait for 1-4 hours and then start cytokine stimulation. Be aware that cells do not like serum-free medium and this change to serum-free conditions may trigger apoptosis; so for e.g. primary endothelial cells (HUVEC) you are limited to 12 hrs serum free conditions bevore apoptosis starts. Good luck!
Hello,
What do you mean by primary cells? Are these from hematopoietic compartment? If so, which lineage? Lymphoid or myeloid?
Pooja, you can perform the treatment with medium containing low serum. I have only used serum starvation (overnight), but if your cells don't do well in serum-free conditions you can reduce your serum to say 0.5-1%.
Best.
Thanks a lot for your suggestions. Hope I could perform my experiment without any difficulty.
I think and cited Yuan Zhai ant authors that is very correct opinion about role cytokines in Organ Transplantation: Th1 and Th2 Cytokines in Organ Transplantation: Paradigm Lost?
Ronald W. Busuttil
Ftafik M. Ghobrial
Jerzy W. Kupiec-Weglinski
The Dumont-UCLA Transplant Center, University of California School of Medicine, 10833 Le Conte Ave, Los Angeles, CA 90095
Yuan Zhai
RESUMO
Identification of CD4+ T helper lymphocyte subsets that exhibit distinct cytokine elaboration patterns has provided a valuable framework for understanding the heterogeneity of the immune response. Much progress has been made in recent years in defining the cellular and molecular mechanisms by which cytokines induce T cell differentiation. In transplantation models, the Thl cytokine profile often associates with allograft rejection, while the Th2 profile favors the acquisition of tolerance. However, this paradigm may not be sufficient to explain the recently demonstratedin vivo effects of cytokine manipulation on allograft survival. Th2 cytokines may not be necessary for tolerance induction, while Thl cytokines may even be beneficial in promoting allograft survival. However, such data should be interpreted in light of the diverse and often redundant effects displayed by cytokine networks in vivo. Understanding the complex interactions of cytokines in the alloimmune cascade therefore is critical for designing therapeutic strategies that abrogate allograft rejection and induce donor-specific tolerance, an elusive goal in organ transplantation.
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