Fig. 5: Cellular fate detection for transplanted BM-MSCs in fibrotic livers.

A Distribution of Luc⁺ BM-MSCs transplanted into fibrotic mice was detected by in vivo imaging system (PerkinElmer). 1 × 10⁶ Luc⁺ BM-MSCs from the male donors were injected into female C57BL/6 J mice 24 h after the last CCl₄ injection. Mice received D-luciferin (150 µg/kg) and were imaged at different times after infusion (n = 3). B Quantification of the luminescence radiance detected by an in vivo imaging system at different times after transplantation (n = 3). C Quantitative real-time PCR analysis for the transcriptional expression of Y chromosome-specific ymt2/b gene to illustrate the number of remaining BM-MSCs in fibrotic livers at different times after transplantation (n = 3). D 1 × 10⁶ Luc⁺ BM-MSCs were injected into C57BL/6 J mice 24 h after the last CCl₄ injection. Mice received Z-DEVD-aminoluciferin (150 µg/kg) and were imaged at 4 h after infusion (n = 3). Z-DEVD-aminoluciferin is a modified firefly luciferase substrate that can be cleaved by caspase-3 to liberate aminoluciferin in apoptotic Luc⁺ BM-MSCs, latter of which is consumed by luciferase to generate a luminescent signal. E 1 × 10⁶ Luc⁺ BM-MSCs were treated by 50% ILCM (v/v) for 4 h, and Z-DEVD-aminoluciferin (50 µg/well) was used to detect apoptosis by an in vivo imaging system. F Representative flow cytometry plots of CM-Dil⁺ cells in the liver. The majority of CM-Dil⁺ signal was identified as Ly6C^lo macrophages. G Proportion of Kupffer cells (F4/80^hiLy6C^lo), infiltrating Ly6C^hi macrophages (F4/80^intLy6C^hi), and Ly6C^lo macrophages (F4/80^intLy6C^lo) in the population of CM-Dil⁺ macrophages (n = 3). Bars = means ± SD; Statistical evaluation was performed using one-way ANOVA with posthoc LSD test; n.s. p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001.