Stem cell therapy for liver disease: parameters governing the success of using bone marrow mesenchymal stem cells

Abstract

Background & aims: Liver transplantation is the primary treatment for various end-stage hepatic diseases but is hindered by the lack of donor organs and by complications associated with rejection and immunosuppression. There is increasing evidence to suggest the bone marrow is a transplantable source of hepatic progenitors. We previously reported that multipotent bone marrow-derived mesenchymal stem cells differentiate into functional hepatocyte-like cells with almost 100% induction frequency under defined conditions, suggesting the potential for clinical applications. The aim of this study was to critically analyze the various parameters governing the success of bone marrow-derived mesenchymal stem cell-based therapy for treatment of liver diseases.

Methods: Lethal fulminant hepatic failure in nonobese diabetic severe combined immunodeficient mice was induced by carbon tetrachloride gavage. Mesenchymal stem cell-derived hepatocytes and mesenchymal stem cells were then intrasplenically or intravenously transplanted at different doses.

Results: Both mesenchymal stem cell-derived hepatocytes and mesenchymal stem cells, transplanted by either intrasplenic or intravenous route, engrafted recipient liver, differentiated into functional hepatocytes, and rescued liver failure. Intravenous transplantation was more effective in rescuing liver failure than intrasplenic transplantation. Moreover, mesenchymal stem cells were more resistant to reactive oxygen species in vitro, reduced oxidative stress in recipient mice, and accelerated repopulation of hepatocytes after liver damage, suggesting a possible role for paracrine effects.

Conclusions: Bone marrow-derived mesenchymal stem cells can effectively rescue experimental liver failure and contribute to liver regeneration and offer a potentially alternative therapy to organ transplantation for treatment of liver diseases.

Conflict of interest statement

The authors report that no conflicts of interest exist.

Figures

Figure 1

MDHs and MSCs rescue lethal FHF. (A) Effect of CCl4 on NOD-SCID mice survival at 0.2–0.4 mL/kg administered by gavage. Survival curves of NOD-SCID mice that underwent intrasplenic transplantation with 1.4 × 106 to 4.2 × 107/kg of (B) MDHs and (C) MSCs and intravenous transplantation with 1.4 × 106 to 4.2 × 107/kg of (D) MDHs and (E) MSCs at 24 hours after administration of 0.28 ml/kg CCl4. Statistical analysis was performed by log-rank test (intrasplenic MSC vs intrasplenic MDH: 4.2 × 107/kg, P = .0001; 1.4 × 107/kg, P = .0018; 4.2 × 106/kg, P = .0190; 1.4 × 106/kg, P = .0190; intravenous MSC vs intravenous MDH: 4.2 × 107/kg, P = .1380; 1.4 × 107/kg, P = .0190; 4.2 × 106/kg, P = .0047; 1.4 × 106/kg, P = .0009).

Figure 2

Transplanted donor cells engraft recipient liver and differentiate into hepatocytes. (A) FACS for human B2M-expressing cells at 4 weeks posttransplantation of MSCs (MSC-Tx). Human B2M-sorted cells (hB2M+) were (B) analyzed for expression of liver marker genes by Western blot and (C) verified for their human origin by genomic DNA polymerase chain reaction. (D) Detection of human albumin in the peripheral blood of recipients rescued by MSC transplantation (enzyme-linked immunosorbent assay, mean ± SD of 3 determinations).

Figure 3

MSCs are more resistant to oxidative stress than MDHs. Effect of exogenous oxidant (A) paraquat and (B) hydrogen peroxide on the viability of MSCs and MDHs (mean ± SD of 3 determinations). (C) Basal expression of antioxidative enzymes in MSCs and MDHs by Western blot. CuZnSOD, copper/zinc superoxide dismutase; MnSOD, manganese superoxide dismutase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. >Change in expression of antiapoptotic genes (D) Bcl-2 and (E) Bcl-xL in MSCs and MDHs in the presence of exogenous oxidant paraquat (mean ± SD of 3 determinations). (F) Changes in reduced glutathione/oxidized glutathione ratio in mice that underwent transplantation with MSCs and MDHs at 24 hours after administration of 0.28 mL CCl4/kg (mean ± SD of 3 determinations). Statistical analysis was performed by paired t test; P < .05 was considered significant. *Statistical significance (D–F).

Figure 4

Transplantation of MSCs rescues endogenous hepatocytes. (A) Mean percentage of necrosis in the liver of recipient mice that underwent transplantation with MSCs and MDHs at 3 and 6 days after CCl4 administration (mean ± SD of 10 determinations). (B) Change in the number of donor-derived cells in the liver of recipient mice that underwent transplantation with MSCs and MDHs at 3, 5, and 7 days after CCl4 administration (mean ± SD of 3 determinations). Bromodeoxyuridine incorporation in the liver of (C) MSC recipients and (D) MDH recipients at 3 and 5 days after CCl4 administration (red arrowheads, QuantumDot-labeled donor cells; blue, nuclei staining; brown nuclei, bromodeoxyuridine-labeled cells). (E) Effect of MSC and MDH coculture on the proliferation of hepatocytes after paraquat treatment (mean ± SD of 3 determinations). Statistical analysis was performed by paired t test; P < .05 was considered significant. *Statistical significance (A,B,E).