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

J Immunol Methods. 2003 Apr 1;275(1-2):251-5. doi: 10.1016/s0022-1759(03)00010-3.


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.

MeSH terms

  • Antibodies, Monoclonal / administration & dosage*
  • CD28 Antigens
  • CD3 Complex
  • Humans
  • Immunologic Techniques*
  • In Vitro Techniques
  • Lymphocyte Activation* / drug effects
  • Microspheres
  • Phytohemagglutinins / pharmacology
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology*


  • Antibodies, Monoclonal
  • CD28 Antigens
  • CD3 Complex
  • Phytohemagglutinins