Efficient Non-Viral Integration and Stable Gene Expression in Multipotent Adult Progenitor Cells
Figure 3
Non-viral integration efficiency in MAPC. Nucleofected rMAPC were plated and the medium was supplemented with G418 (400 g/mL) one day later. (a) Colony-forming assay. After 10–12 days of growth under selective conditions, cells were fixed and stained to determine the frequency of G418-resistant colony formation. The number of G418-resistant colonies () for each group is shown ± S.D. (b) Cells were harvested into a suspension and viable cell counts were performed by trypan blue exclusion on the indicated days and the total number of cells in culture is reported as mean ± SE. (c) Genomic DNA isolated from the individual clones obtained with SB and Int was subjected to restriction enzyme digestion, plasmid sequence rescue, and sequencing of the recovered fragment carried out to determine the genomic site of integration.
