Figure 2.

Enhanced Engraftment of HuSCs by Multiple-Site Injection

(A) Representative images of a conventional transplant with a single injection with 10,000 cells per TA (n = 5, biological replicates). TA cross-sections were stained with human DYSTROPHIN (left) for human fibers or with human-specific SPECTRIN, LAMIN A/C, Laminin, and PAX7 for HuSCs (right). Satellite cells are marked with an arrow.

(B) Bar graph representation of human fiber engraftment with varying dosage of transplanted cells. Isolated satellite cells were transplanted in a single injection of either 1,000 (n = 3), 5,000 (n = 9), or 10,000 (n = 5) cells. n Values denote biological replicates. Human DYSTROPHIN-positive fibers were counted in TA cross-sections, and the y axis value indicates the number of fibers within the cross-section containing the maximum number of human-derived fibers. The number of engrafted fibers was not significantly different among the three groups.

(C) Satellite cells were transplanted in either a single injection (top panels) or with multiple injection sites (bottom panels) with a dose of 2,000 cells per TA. TA cross-sections were stained with human DYSTROPHIN (left) for human fibers or with human-specific SPECTRIN, LAMIN A/C, Laminin, and PAX7 for HuSCs (right) (n = 4 biological replicates). Satellite cells are marked with arrows. Scale bars, 100 μm (left panels; also applies to A) and 10 μm (right panels; also applies to A).

(D) Bar graph showing the engraftment of human myofibers after transplantation as assessed by human DYSTROPHIN staining (n = 4 per group, individual mice).

(E) Bar graph showing the average engraftment of PAX7-positive HuSCs per cross-section after transplantation (n = 4 per group, individual mice).

Data presented as mean ± SEM. ^∗p < 0.05. All samples were processed the morning after tissue collection, within 12 hr after muscle biopsy. All mice were analyzed 5 weeks after transplantation. See also Figure S3.