I am new to immunology and T cell biology. I know that in vivo T cells have different subsets based on CD44 and CD62 expression, e.g. memory T cells express CD44. How about in vitro cultured T cells? Mice spleen CD3 T cells are isolated by Pan T cell isolation kit (negative selection), and activated for 48 hour via plate-bound CD3/CD28 antibodies (2ug/ml and 1ug/ml, respectively). Then the cells are transferred to uncoated plate and maintained in culture (RPMI medium, with 10% FBS, 30ng/ml IL-2, P/S, 2-BME, non-essential AA). How will the cells differentiate by CD44 and CD62 expression? Thanks a lot.
I have only checked CD62L and CD44 expression on day 3 or day 4 (2 days after removal from anti-CD3 and -CD28 coated wells) and the cells were CD62L+ CD44+. Have not checked them at later stages of cell culture.
From my result, CD44 will be upregulated within 24 hours of CD3/CD28 stimulation and remained high expression for at least 5 days in cell culture. I did not culture longer than 5 days, so not result can be shared.
Once stimulated, T cells are supposed to downregulate CD62L and upregulate CD44 as part of becoming activated.
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