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Effects of different cell culture medium and different sources of serum on proliferative and functional activities of cytokine-induced killer cells
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Abstract:
Objective:To investigate the effects of different culture media and different sources of serum on proliferative and functional activities of cytokine-induced killer (CIK) cells. Methods: Mononuclear cells were isolated from peripheral blood of 6 lung cancer patients and were induced to differentiate into CIK cells in 8 different culture conditions: GT-T551+autologous serum, GT-T551 + serum from a healthy individual, GT-T551 + FBS, GT-T551, RPMI 1640 + autologous, RPMI 1640 + serum from a healthy individual, RPMI 1640 + FBS, and RPMI 1640. After induction for 6 days, cell proliferation was assessed carboxyfluorescein diacetate succinimidyl sster (CFSE) labeling assay, and CD3+CD8+ cell proportion, CD3+CD4+ T cell expression of Granzyme B and IFN-γ and perforin and the expression of CD107a on CD3+CD56+, CD3+CD4+ and CD3+CD8+ T cells in the presence of leukemia NB4 and K562 cells as targets were assessed by flow cytometry. Results: The proliferation index of CIK cells cultured in GT-T551 medium supplemented with autologous serum was significantly higher than that in other culture conditions ( P <0.05). The expression of Granzyme B on CD4+ T cells was significantly higher ( P <0.01) in the GT-T551+autologous serum group \[22.85±3.50\]% than in the GT-T551+healthy serum group \[13.28±1.75\]% and significantly higher ( P <0.01) in the RPMI 1640+autologous serum group \[22.57±3.45\]% than in the RPMI 1640+healthy serum group\[15.37±4.08\]%. Furthermore, the amount of IFN-γ secreted from CD8+ T cells was significantly higher in the GT-T551 + autologous serum groups compared to GT-T551+healthy serum group. In the presence of killing NB4 and K562 cells, the expression of CD107a on the CD3+CD56+NKT cells was significantly higher in the GT-T551+autologous serum group (\[7.10±1.94\]%, \[8.00±1.82\]%) than in the GT-T551+healthy serum group (\[2.73±0.79\]%, \[3.03±0.78)%; P <0.01) and in the RPMI 1640+autologous serum group (\[4.45±1.96\]%, \[3.30±1.47\]%; all P <0.01). Conclusion: GT-T551 medium supplemented with autologous serum appears the optimal in vitro system to stimulate proliferation, cytokine secretion and enhance the cytotoxicity of CIK cells. This finding may have some clinical significance in CIK therapy.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81171986，No. 81271815), the Research Grant from the Chinese Ministry of Health (No. 20110110001), the Basic and Advanced Technology Research Foundation from Science and Technology Bureau of He’nan Province (No.112300410153, No.122300410155), the Program of Innovation Talents of Science and Technology from Science and Technology Burea u of He’nan Province (No.124200510006), and the Foundation for Innovation Team of the First Affiliated Hospital of Zhengzhou University
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