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J Immunol Methods. 2003 Apr 1;275(1-2):251-5.

T cell stimulation and expansion using anti-CD3/CD28 beads.

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  • 1Clinical Haematology and Haematology-SEALS, St. George Hospital, Sydney, NSW, Australia.


Following appropriate stimulation, T lymphocytes will proliferate extensively in vitro. Traditionally, mitogenic lectins such as phytohemagglutinin (PHA) and concanavalin A (Con A) have been used for polyclonal T cell stimulation. A more physiologically relevant approach uses beads coated with anti-CD3 and anti-CD28 to stimulate T cells in a manner that partially mimics stimulation by antigen-presenting cells. This protocol describes the steps involved in T cell stimulation and their subsequent in vitro expansion using anti-CD3/CD28 beads.

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