Hi, scholars.
I am planning to conduct mouse neutrophil and T cell co-culture in Vitro and observe the effect of neutrophils on T cells.
I don't have any protocol. Could anyone provide me protocol and the reagents such as do I need to add any recombinant cytokines or peptide or anti-CD3 and CD28 to the culture medium. How long should I culture the T cells and neutrophils to T cell ratio.
Thank you.
The protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans, it also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence-activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. This protocol described in :
Isolation of mouse neutrophils
Muthulekha Swamydas, Yi Luo, Martin E Dorf, Michail S Lionakis
Current protocols in immunology 110 (1), 3.20. 1-3.20. 15, 2015
Thank you so much for your recommendations.
After isolating neutrophils, I would like to know know how to set up neutrophil and T cells co-culture methods. Do we need to add any recombinant cytokines, chemokines or Anti-CD3 and CD28 to the Neutrophil- T cell co-culture and how long should we stimulate and other parameters.
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