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Fig. 4 NFA and LPA treatments enhance the engraftment efficiency of mouse satellite cells (A) Experimental design. (B and C) Engraftment efficiency of 4-h-treated mouse satellite cells with vehicle, LPA (5 μM), or NFA (10 μM). Five thousand treated cells from FVB-Tg(CAG-eGFP) mice were injected into pre-injured TA muscles of non-transgenic FVB/NJ recipient, and engraftment was measured by counting the number of GFP+ myofibers. Data are presented as mean ± SEM. Statistical analysis by unpaired t test; both treatment groups were significantly different from the control group (n = 10). (D) Transverse frozen section of TA muscles injected with 5,000 treated satellite cells. Cell membrane (WGA, red), nuclei (DAPI, blue), and engrafted cells (GFP, green). These studies used 8- to 16-week-old male mice as recipients. DMSO concentration for all groups including experimental control and as vehicle for NFA and LPA was 0.1%. Scale bar, 200 μm. (E) Engraftment efficiency of 4-h-treated mouse satellite cells with vehicle, LPA (5 μM), or NFA (10 μM). Two thousand treated cells from C57BL/CAG-EGFP mice were injected into pre-injured TA muscles of PrkdcscidDmdmdx recipients, and engraftment was measured by counting the number of GFP+ myofibers. Data are presented as mean ± SEM. Statistical analysis by one-way ANOVA; both treatment groups were significantly different from the control group (n = 9 for vehicle and n = 7 for LPA and NFA treatment groups). (F) Restoration of dystrophin in engrafted PrkdcscidDmdmdx recipients using a mouse-specific dystrophin antibody (Sigma, D8043). Nuclei (DAPI, blue), engrafted cells (GFP, green), and dystrophin (Alexa Fluor 594, red). Scale bar, 50 μm. (G) NFA and LPA treatments do not change the rate of incorporation of transplanted satellite cells into the in vivo pool of muscle precursors. Donor mouse satellite cells were isolated from Pax7:ZsGreen mice and treated for 4 h with vehicle (0.1% DMSO), LPA (5 μM), or NFA (10 μM). Five thousand treated cells were transplanted into pre-injured TA muscles of WT (C57BL/6J) recipients, and the engrafted donor satellite cells (ZsGreen+) were counted via flow cytometry 4 weeks after transplantation. Data are presented as mean ± SEM. Statistical analysis by one-way ANOVA with Dunnett’s correction (n = 4 for vehicle and n = 5 for LPA and NFA treatment groups). These studies used adult (8–16 weeks of age) male mice.