Human Liver-Derived Stem Cells Improve Fibrosis and Inflammation Associated with Nonalcoholic Steatohepatitis

Figure 5

Effects of HLSCs on liver morphology and fibrosis. (a) Representative light microscopy micrographs of liver histology of mice at week 4 of MCDD, treated with intravenous injection of HLSCs or with vehicle alone (NASH) (original magnification 400x). Sirius Red staining showed fibrosis in the perivenous area of MCDD-fed mice. Red stain represents collagen fibers considered to be a marker of liver fibrosis. (b) Histological quantification of fibrosis in MCDD-fed mice injected with HLSCs or with vehicle alone (NASH) at different weeks, by multiphase image analyses of 10 fields per section (original magnification 400x). Data shown represents . ANOVA with Newman-Keuls multicomparison test was performed: MCDD-fed mice injected with HLSCs at different weeks (1 , 2 , or 3 ) versus MCDD-fed mice injected with vehicle alone (NASH). (c) Histological quantification of the surface area occupied by steatosis vacuoles in MCDD-fed mice injected with HLSCs or with vehicle alone (NASH) at different weeks (1 , 2 , or 3 ) by multiphase image analyses of 10 fields per section (original magnification 200x). Data shown represents . (d) Quantification of PCNA-positive cells/high-power field (HPF) (original magnification ×400) in 10 fields per section. Data represents . ANOVA with the Newman-Keuls multicomparison test was performed: MCDD-fed mice injected with HLSCs at different weeks (1 , 2 , or 3 ) versus MCDD-fed mice injected with vehicle alone (NASH). (e) Gene expression levels of fibrotic markers Tgf-β, Col 1a1, and α-Sma in livers of MCDD-fed mice treated with vehicle alone or with HLSCs () at different weeks (1 , 2 , or 3 ). Data are expressed as relative quantification using the ΔΔCt method. Normalization was made using GAPDH as a housekeeping gene. ANOVA with the Newman-Keuls multicomparison test was performed; , , and HLSC-treated mice versus NASH mice treated with vehicle alone; MCDD-fed mice NASH treated with vehicle alone versus control animals fed with standard diet.

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