title: Impact of culture medium on the expansion of T cells for immunotherapy.
abstract: OBJECTIVE
  Encouraging evidence of clinical benefits from cancer immunotherapy is beginning to accumulate in several clinical trials. Cancer immunotherapy is based on two main methods, active vaccination and cell-transfer therapy. The ex vivo expansion of T cells is required to monitor vaccine-induced antigen-specific T cells or prepare large numbers of reactive lymphocytes for adoptive transfer.
  
  
  METHODS
  We examined the influence of culture medium on T-cell growth, cytotoxicity and phenotype after activation using immobilized anti-CD3 monoclonal antibody or Zoledronate stimulation. Peripheral blood mononuclear cells (PBMC) were cultured in RPMI, AIM-V or OpTmizer with or without autologous serum.
  
  
  RESULTS
  When supplemented with sufficient serum, RPMI was a good culture medium for T-cell expansion following anti-CD3 stimulation. Addition of autologous serum to AIM-V or OpTmizer increased the numbers of cells obtained to a similar extent, but their phenotype and function were quite different. Activated T cells cultured with OpTmizer mediated greater cytotoxicity than any other culture. Regardless of the media used, the main population expanded after CD3 stimulation was CD3(+) CD8(+). While more CD3(+) CD4(+) T cells were induced in RPMI and AIM-V, more CD3(-) CD56(+) cells and CD3(+) CD56(+) T cells were induced in OpTmizer. When cells were stimulated by Zoledronate for 14 days, approximately 7.2 times and 11.5 times more gammadelta T cells were obtained in OpTmizer than AIM-V or RPMI, respectively.
  
  
  CONCLUSIONS
  Successful immunotherapy depends on the selection of appropriate culture media to support efficient expansion of the type of T cell desired.
authors.affiliation: Department of Immunotherapeutics (Medinet), Graduate School of Medicine, The University of Tokyo, Bunkyo-Ku, Tokyo, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Sato
authors.forename: Makoto
authors.initials: M
authors.lastname: Kondo
authors.forename: Kazuko
authors.initials: K
authors.lastname: Sakuta
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hosoi
authors.forename: Shuichi
authors.initials: S
authors.lastname: Noji
authors.forename: Miki
authors.initials: M
authors.lastname: Sugiura
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Yoshida
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Kakimi
mesh_terms: D000911:Antibodies, Monoclonal; D017252:CD3 Complex / Q000276:immunology; D049109:Cell Proliferation; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism*; D003602:Cytotoxicity, Immunologic; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008213:Lymphocyte Activation; D009369:Neoplasms / Q000276:immunology / Q000628:therapy*; D044967:Serum / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism* / Q000473:pathology
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D017077:Culture Media, Conditioned