Fig. 4

Human muscle stem cells (hMuSCs) alleviate acute liver injury. (A) PBMCs stained with CFSE were co-cultured with hMuSCs (2.5 × 10⁴ cells per well in 48-well plates) at graded ratios (hMuSC : PBMC) for 72 h in the presence of anti-human CD3 antibody (1 µg/ml). CFSE fluorescence intensity reduction of PBMCs was detected by flow cytometry. (B) The proliferation of PBMCs was measured by the CFSE dilution assay (n = 4). (C) Mice were intravenously injected with ConA (15 mg/kg) to induce acute liver injury; 0.5 h later, hMuSCs (5 × 10⁵) stimulated with human IFN-γ and TNF-α (10 ng/ml each) were transfused to suffering mice in the same way. After 12 h, serum and liver tissues were sampled. (D) Serum levels of AST and ALT were measured. (E) IFN-γ and IL-12 levels in the serum were assayed by ELISA. (F) Representative H&E-stained liver sections and percentages of tissue necrosis. Scale bar, 250 μm. (G) Absolute numbers of MNCs in liver tissues were calculated. Absolute numbers of CD3⁺ T cells were determined by flow cytometry (Control: n = 5, ConA + PBS: n = 5, ConA + hMuSC: n = 5). Values are presented as mean ± SEM. Statistical analysis was performed by Mann-Whitney test.