Expansion of α-galactosylceramide-stimulated Vα24+ NKT cells cultured in the absence of animal materials

Yukie Harada, Osamu Imataki, Yuji Heike, Hiroyuki Kawai, Akihiro Shimosaka, Shin Ichiro Mori, Masahiro Kami, Ryuji Tanosaki, Yoshinori Ikarashi, Akira Iizuka, Mitsuji Yoshida, Hiro Wakasugi, Shigeru Saito, Yoichi Takaue, Masao Takei, Tadao Kakizoe

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)


Vα24+ NKT is an innate lymphocyte with potential antitumor activity. Clinical applications of Vα24+ natural killer (NK) T cells, which are innate lymphocytes with potential antitumor activity, require their in vitro expansion. To avoid the potential dangers posed to patients by fetal bovine serum (FBS), the authors evaluated non-FBS culture conditions for the selective and efficient expansion of human Vα24+ NKT cells. Mononuclear cells (MNCs) and plasma from the peripheral blood of normal healthy donors were used before and after G-CSF mobilization. MNCs and plasma separated from apheresis products were also used. MNCs were cultured for 12 days in AIM-V medium containing α-galactosylceramide (α-GalCer) (100 ng/mL) and IL-2 (100 U/mL) supplemented with FBS, autologous plasma, or autologous serum. The cultured cells were collected and their surface markers, intracellular cytokines, and cytotoxicity were evaluated. The highest expansion ratio for Vα24+ NKT cells was obtained from G-CSF-mobilized MNCs cultured in medium containing 5% autologous plasma. Cultures containing MNCs and autologous plasma obtained before and after G-CSF mobilization had approximately 350-fold and 2,000-fold expansion ratios, respectively. These results suggest that G-CSF mobilization conferred a proliferative advantage to Vα24 + NKT cells by modifying the biology of cells and plasma factors. Expanded Vα24+ NKT cells retained their surface antigen expression and production of IFN-γ and exhibited CD1d-independent cytotoxicity against tumor cells. Vα24+ NKT cells can be efficiently expanded from G-CSF-mobilized peripheral blood MNCs in non-FBS culture conditions with α-GalCer and IL-2.

Original languageEnglish
Pages (from-to)314-321
Number of pages8
JournalJournal of Immunotherapy
Issue number4
Publication statusPublished - 2005
Externally publishedYes


  • G-CSF
  • NKT cells
  • α-galactosylceramide

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology
  • Cancer Research


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