Ex Vivo Expansion of Murine MSC Impairs Transcription Factor-Induced Differentiation into Pancreatic β-Cells

Figure 2

NOD-derived MSC nucleofection. (a) MSCs (early passage number) were nucleofected with 0 and 5 μg pVITRO2-Luc2. Parental MSCs and MSC-Luc2 at an equivalent passage number (P15) showed native fibroblast-like morphology and maintained plastic adherence and self-renewal properties. Images were acquired on a Leica DM light microscope at 10x magnification, scale bar = 100 μm. (b) In vitro functional characterization of luciferase activity in MSC-Luc2. Cells were incubated with 1 : 1 D-luciferin (300 μg/ml) and imaged on the IVIS Lumina II, according to the in vitro BLI acquisition settings. The image represented is at  min after the addition of D-luciferin. Lane 1: D-PBS; lanes 2-10: MSC-Luc2 and MSC-Luc2/LacZ (control). (c) Linear regression analysis of luminescent signal was performed using GraphPad Prism 7^®. Data are presented as means ± SDs of triplicates.

(a)

(b)

(c)