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A Comparison of Two Methods to Expand Regulatory T Cells (Tregs) In Vitro
Liu Zhao, Luo Shiqiao*
Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
Abstract: CD4+CD25+Tregs (regulatory T cells) and CD4+T cells were isolated from BALB/c spleens by MACS (immunomagnetic beads)， respectively. CD4+CD25+Tregs were cultured for 14 d in 96-well plates coated with anti-CD3/CD28 microbeads in the presence of 1 000 U/mL of IL-2; then CD4+T cells were cultured in 24-well plates coated with anti-CD3/CD28 microbeads in the presence of 100 U/mL IL-2 and 5 ng/mL of recombinant human TGF-β (transforming growth factor-β) for 5 d. The immunosuppression and counts of Tregs expanded by the two methods were analyzed. The experiment results showed that there was no obvious difference in immunosuppression between the two methods of expanding Tregs in vitro (P>0.05)， but when used the same size BALB/c mice， we can get (4.9±0.4)×107 Tregs through CD4+T cells and only get (1.9±0.1)×107 Tregs through CD4+CD25+Tregs (P<0.05). The result suggested that there was no obvious difference in immunosuppression between the two methods of expanding Tregs， but get more Tregs and take less time through CD4+T cells.
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