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. Author manuscript; available in PMC: 2019 May 19.
Published in final edited form as: Gene Ther. 2011 May 5;18(11):1078–1086. doi: 10.1038/gt.2011.63

Figure 1.

Figure 1

Effects of additional serum or IL-3 on human CD34+ cell transduction. (a) As in our basic condition, we cultured human CD34+ cells with cytokines alone for 24h (prestimulation), with cytokines and a GFP-expressing HIV1-based lentiviral vector for an additional 24h (transduction). (b) To evaluate whether FBS has effects on human CD34+ cell transduction, we performed transduction at an MOI of 5 in DMEM with 10% FBS or serum-free X-VIVO10 medium, and we analyzed GFP expression rates (%GFP), mean fluorescence intensity (MFI) of GFP and CD34+ rates in bulk cells (%CD34). (c) To evaluate the effects of FBS at high MOI, we transduced human CD34+ cells at an MOI of 50, and analyzed %GFP, MFI of GFP and cell viability (Trypan blue stain). (d) To evaluate effects of supplemental IL-3 to the 100 ng ml−1 cytokine mixture (Mix) in serum-free X-VIVO10 medium, we compared Mix, addition of 20 ng ml−1 IL-3 (+IL-3) and addition of 60 ng ml−1 IL-3 (3×IL-3).