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. Author manuscript; available in PMC: 2016 Jul 18.
Published in final edited form as: Cytotherapy. 2014 Sep;16(9):1257–1269. doi: 10.1016/j.jcyt.2014.05.022

Figure 1.

Figure 1

Generation of piggyBac-mediated CD19-specific T cells in serum/xeno/tumor cell/virus-free culture. Mononuclear cells from 10 mL of blood of healthy donors were nucleofected with pIRII-CAR.CD19-28-ξ and pCMV-piggyBac with the use of the 4D-Nucleofector Device and immediately transferred into IL-15–containing TexMACS medium. On day 1, nucleofected cells were stimulated with the use of CD3/CD28 mAbs for 4 days. Six days after the stimulation, cells were selected with the use of an anti-CAR antibody and restimulated with CD3/CD28 mAbs for 4 days, then cultured for another 10 days in IL-15–containing TexMACS with autologous feeder cells. Twenty-one days after the gene transfer, cells were harvested, analyzed and cryopreserved. Data are presented as mean ± standard deviation of experiments from three donors.