Fig. 2

Delivery of Multiple DNA Vectors in hMSCs for Expression of Two Reporter Transgenes.  hMSCs were transfected with single transgene vectors (pEGFP or ptdTomato) complexed with Lipofectamine 3000 (a, c, e, & g) or Turbofect (b, d, f, & h) and transfection efficiencies [i.e., number of EGFP (green bars), tdTomato (red bars), and co-expressing (both EGFP and tdTomato expression, yellow bars) cells relative to total cell counts] for each transgene (EGFP or tdTomato) were compared to two transgene delivery strategies; (i) two DNA vectors delivered as separate complexes ([pE]+[pT]); and (ii) two DNA vectors delivered in the same complex ([pE + pT]). Transfection efficiencies for hMSCs expressing both EGFP and tdTomato (co-expression) was calculated by dividing the number of cells that were both EGFP and tdTomato positive by the total cell count (Hoechst, nuclear stain). a Transfection efficiencies for all conditions in D1 hAMSCs that used Lipofectamine 3000 as the cationic transfection reagent. b Transfection efficiencies for all conditions in D1 hAMSCs that made use of Turbofect as the cationic transfection reagent. c Transfection efficiencies for all conditions in D2 hAMSCs that used Lipofectamine 3000 as the cationic transfection reagent. d Transfection efficiencies for all conditions in D2 hAMSCs that made use of Turbofect as the cationic transfection reagent. e Transfection efficiencies for all conditions in D3 hBMSCs that used Lipofectamine 3000 as the cationic transfection reagent. f Transfection efficiencies for all conditions in D3 hBMSCs that made use of Turbofect as the cationic transfection reagent. g Transfection efficiencies for all conditions in D4 hBMSCs that used Lipofectamine 3000 as the cationic transfection reagent. h Transfection efficiencies for all conditions in D4 hBMSCs that made use of Turbofect as the cationic transfection reagent. All conditions have equal moles of expression cassette and mass of DNA delivered. Data represented as mean ± SEM (n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant (p > 0.05), as determined by a 2-way ANOVA with Tukey’s post hoc test