Bone marrow-derived mesenchymal stem cells protect against experimental liver fibrosis in rats

Figure 1 MSCs are induced to differentiate into osteoblasts and stained red color by Chinalizarin (A). Under adipogenic conditions, MSCs accumulate lipid vacuoles, which are positively stained by Oil Red O assay (B). Under neurogenic conditions, MSCs differentiate into neuronal-like cells and stained positively for NSE by immunocytochemical analysis (C). (Original magnification, ×200.)

Figure 2 Average weekly weight of rats induced with CCl₄ (A) or DMN (B). A: Paraffin/saline, n = 10; CCl₄/saline, n = 10-8; CCl₄/MSCs, n = 10-9; B: saline, n = 10; DMN/saline, n = 40-4; DMN10/MSCs, n = 10-8; DMN20/MSCs, n = 10-6.

Figure 3 Analysis of liver histopathology and immunohistochemistry in rats induced with CCl₄. A: Paraffin/saline; B: CCl₄×6 wk; C: CCl₄/MSCs; D: CCl₄/saline; 1: HE staining, original magnification ×100; 2: MT staining, original magnification ×40; 3: immunohistochemistry for α-SMA, DAB staining, original magnification ×400; E: MT and α-SMA staining was quantified using IBAS 2.5 software. Data represent the fold-increase in positive staining vs paraffin/saline. Values are presented as mean±SD. ^bP<0.01 vs CCl₄/saline. Not significantly different between values of CCl₄/saline and CCl₄×6 wk.

Figure 4 Survival of rats in control group and DMN-induced group with or without MSCs administration. Analysis of survival was conducted by a log-rank test based on the Kaplan–Meier method.

Figure 5 Analysis of liver histopathology and immunohistochemistry in rats induced with DMN. A: DMN10; B: DMN20; C: DMN10/MSCs; D: DMN20/MSCs; E: DMN10/saline; 1: HE staining, original magnification ×100; 2: MT staining, original magnification ×100; 3: immunohistochemistry for α-SMA, DAB staining, original magnification ×400; F: MT and α-SMA staining was quantified using IBAS 2.5 software. Data represent the fold-increase in positive staining vs saline. Values are presented as mean±SD. ^aP<0.05 vs DMN20/MSCs, ^bP<0.01 vs DMN10/saline respectively.

Figure 6 PCR signals of sry gene and β-actin gene in liver of rats. sry, 104 bp; β-actin, 206 bp; M: 100-bp DNA maker; lane 1: CCl₄/MSCs; lane 2: CCl₄×6 wk; lane 3: DMN10/saline; lane 4: DMN10/MSCs; lane 5: DMN20/MSCs; lane 6: a male Fischer rat 344 as positive control; lane 7: a female Wistar rat as negative control.

Figure 7 Signals of SRY by FISH with Rat 12/Y whole chromosome probes. Red color represent chromosome Y, green color represent chromosome 12, blue color represent nuclear. Original magnification ×40.

Figure 8 Morphology of hepatic stellate cells co-culture with or without mesenchymal stem cells by transwell in vitro for 72 h. A: Hepatic stellate cells co-culture with mesenchymal stem cells; B: hepatic stellate cells co-culture with hepatoceyte cells line; C: hepatic stellate cells only culture. Original magnification ×10.

Figure 9 Analysis of hepatic stellate cells by FACS. A: Hepatic stellate cells co-culture with mesenchymal stem cells; B: hepatic stellate cells only culture.