Fig. 1.

Confirmation of ex vivo creation of two Dystrophin Expressing Chimeric (DEC) human cell lines derived from myoblasts of two normal donors (MB^N1/MB^N2) and from myoblasts of normal and DMD donors (MB^N/MB^DMD). a) Diagram of ex vivo polyethylene glycol (PEG) mediated cell fusion procedure to create DEC; left panel, normal myoblast fusion (MB^N1/MB^N2), right panel, fusion of myoblast from normal and DMD-affected donor (MB^N/MB^DMD). (b) Representative immunofluorescence images of MB^N1, MB^N (green), and MB^N2, MB^DMD (red) parent cells before fusion, and MB^N1/MB^N2 DEC and MB^N/MB^DMD DEC after fusion; For merge: Green: PKH67; Red: PKH26; Blue: DAPI (nuclei); Magnification 630X, scale bars 10 µm. (c) Fusion of the MB^N1 and MB^N2 parent cells (top) and MB^N and MB^DMD parent cells (bottom) assessed by FACS. The overlapping fluorescence of PKH67/PKH26 confirms chimeric state for MB^N1/MB^N1 DEC cell line (top - far right) and MB^N/MB^DMD DEC cell line (bottom - far right). (d) PCR-rSSOP analysis of DEC for the presence of parent cell specific alleles from both donors. (e) STR-PCR analysis of DEC for the presence of parent cell specific loci from both donors