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Updated: Aug 29, 2026
Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus
Published on: May 7, 2012
[The experimental study on inducing and expanding T/NK cells from mononuclear cells of human umbilical cord blood]
Ge-yu Chen1, Shao-liang Huang, Dun-hua Zhou
1The Second Affiliated Hospital of Zhongshan University, Guangzhou 510120, China.
Objective:
To explore the most efficient culture system which can induce cord blood (CB)-mononuclear cells (MNC) to differentiate into mature T/NK cells in vitro.
Methods:
The CB MNCs were cultured in six culture systems respectively for 4 weeks. The T/NK cell surface phenotypes were analyzed by flow cytometry and the absolute numbers of nucleated cells (NCs) were counted at each time point. Moreover, cell morphology was identified by Giemsa-Wright staining, and cytotoxicity of the cultured cells to K562 and Raji tumor cells was also evaluated by MTT method.
Results:
Cultured in the cytokine cocktail of SCF + FLT-3L + IL-7 + IL-15 + TNF-alpha + IL-2, the NCs were (20 approximately 26) x 10(6)/ml in numbers at day 22. The percentage of lymphocytes in the NCs and that of CD(3)(+) T cells in the lymphocytes both exceeded 90% at the same time. Most of the CD(3)(+) T cells were CD(3)(+)CD(8)(+) and the percentage of CD(3)(+)CD(4)(+) T cells declined gradually. The percentage of CD(3)(+)CD(56)(+) NKT cells and gamma delta(+)T cells in the lymphocytes arised from lower than 2% to 30% approximately 40% and 10% approximately 15%, respectively. CD(3)(-)CD(56)(+) NK cells were not expanded. The cytotoxic activity of the cultured cells to K562 and Raji cells at an effector:target (E:T) ratio of 50:1 was over 75% and about 32% approximately 65%, respectively.
Conclusion:
The most efficient culture system which can induce CB MNC to differentiate into mature T/NK cells in vitro is the cytokines cocktail of SCF + FLT-3L + IL-7 + IL-15 + TNF-alpha + IL-2, and the optimum culture time is 22 days.
