Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression

Figure 2

Effect of donor variability on hBM-MSC cytotoxicity, viability, surface marker profiling, and efficiency after transfection. (a) Representative 20X fluorescent images of GFP-transfected cells using TransIT-2020 where green represents GFP+ cells. Transfection was done 24 hours prior on 5 different hBM-MSC cultures (12RB, 37RB, 48RB, 56RB, and 85RB). Scale bars represent 200 μm. (b) Quantification by flow cytometry of percent GFP+ cells of 5 hBM-MSC donors. (c) Percentage of viable cells quantified by flow cytometry using SYTOX Orange. (d) Percentage of cytotoxicity in the cells following transfection via quantification of lactate dehydrogenase (LDH). (e) Percentage of cells positive for hBM-MSC CD73, CD90, and CD105 surface markers after transfection. Results from 4 (a, b, c, and e) or 3 (d) independent experiments with technical duplicates, where error bars represent S.E.M. Statistical significance was obtained using multiple t-tests followed by a Holm-Sidak correction. , , and .

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