2B,C). Treatment with intravenous injections of HLSCs led to a significant reduction of apoptosis and necrosis in surviving mice (Fig. 2A), despite the increase in liver enzymes at 7 hours. In mice surviving to injury, a significant decrease of liver enzymes was observed 3 days after HLSC injection, subsequently reaching normal values (Fig. 2B,C). www.selleckchem.com/products/LY294002.html A significantly lower concentration of ammonium was detected in serum of FLF mice (GalN/LPS) treated with HLSCs compared to vehicle alone (Fig. 2D). In HLSC-treated mice, normal liver morphology

was reestablished after 7 to 21 days of treatment (not shown). As shown in Fig. 3, HLSCs significantly inhibited liver apoptosis in FLF mice (GalN/LPS) compared to vehicle alone. Proliferating cell nuclear antigen (PCNA)-positive cells detected at 7 hours expressed human leukocyte antigen (HLA) or CFSE, indicating that they were derived from the injected HLSCs (Fig. 3C). However, after 3 days in mice

treated with HLSCs, PCNA-positive cells were mainly negative for HLA and CFSE, indicating that most proliferating cells were of murine origin (Fig. 3C). Liver cell localization was evaluated by IVIS using DiD-labeled HLSCs.12, 13 As shown in Fig. 4A,B, after intravenous injection HLSCs preferentially accumulated in livers of mice with FLF but not in livers of healthy mice (Fig. 4A,B). Fluorescence signals, expressed as average radiance, increased until day 7 in livers Chloroambucil of mice with FLF but not in those of healthy mice. In LP-injected mice, no difference in liver accumulation www.selleckchem.com/products/FK-506-(Tacrolimus).html of HLSCs between FLF and healthy mice was observed (Supporting Fig. 1S,B). By immunohistochemistry, CFSE-labeled HLSCs were mainly

detected in large liver vessel walls after 24 hours and within the liver parenchyma after days 7 and 21 in intravenously injected surviving FLF mice (Fig. 5A,D,E). In these mice, CFSE-labeled HLSCs were transiently detected after 24 hours in lungs and spleens, decreasing thereafter (Fig. 5B,C; Fig. 2S). When HLSCs were intravenously injected in healthy mice treated with vehicle alone, liver accumulation was significantly lower than in FLF mice (Fig. 5A). In LP-injected mice, CFSE-labeled HLSCs were detected in the liver parenchyma at days 7 and 21 following injection (Fig. 1S,B), but there was no accumulation in lungs or spleens at any timepoint (not shown). To assess whether HLSCs engrafted in the liver expressed mature hepatic markers, we investigated coexpression of human antigen HLA and mature hepatic markers such as cytokeratin 8 and 18 by confocal analysis (Fig. 6). At day 7 the majority of HLA-positive cells expressed cytokeratin 8 and 18 (Fig. 6A,B). At day 21, ∼50% of HLA-positive cells expressed cytokeratin 8/18 (Fig. 6A,B).