Functional integration of hepatocytes derived from human mesenchymal stem cells into mouse livers

Aims: At present, clinical success of hepatocyte transplantation as an alternative to whole liver transplantation is hampered by the limited availability of suitable donor organs for the isolation of transplantable hepatocytes. Hence, novel cell sources are required to deliver hepatocytes of adequate quality for clinical use. Mesenchymal stem cells (MSCs) from human bone marrow may have the potential to differentiate into hepatocytes in vitro and in vivo. Methods: Isolated MSCs were selected by density gradient centrifugation and plastic adherence, differentiated in the presence of human hepatocyte growth medium and transplanted in immunodeficient Pfp/Rag2 mice. Results: Here, we demonstrate that human MSCs gain in vitro the characteristic morphology and function of hepatocytes in response to specified growth factors. Specifically, preconditioned MSCs store glycogen, synthesise urea and feature the active hepatocyte-specific gene promoter of phosphoenolpyruvate carboxykinase (PCK1). After transplantation into livers of immunodeficient mice, preconditioned MSCs engraft predominantly in the periportal portion of the liver lobule. In situ, the cells continue to store glycogen and express PCK1, connexin32, albumin and the human hepatocyte-specific antigen HepPar1, indicating that the transplanted cells retain prominent qualities of hepatocytes after their regional integration. Conclusion: MSCs derived from human bone marrow may serve as a novel source for the propagation of hepatocyte-like cells suitable for cell therapy in liver diseases.

[1]  T. Takayama,et al.  Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion. , 2005, Blood.

[2]  D. Prockop,et al.  Non-hematopoietic bone marrow stem cells: molecular control of expansion and differentiation. , 2005, Experimental cell research.

[3]  S. Hwang,et al.  In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells. , 2005, Biochemical and biophysical research communications.

[4]  李亚明,et al.  Hepatocyte transplantation , 2005 .

[5]  Min-Jeong Seo,et al.  Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. , 2005, Biochemical and biophysical research communications.

[6]  H. Gilgenkrantz.  Mesenchymal stem cells: An alternative source of hepatocytes? , 2004, Hepatology.

[7]  O. Lee,et al.  In vitro hepatic differentiation of human mesenchymal stem cells , 2004, Hepatology.

[8]  M. Kassem,et al.  Mesenchymal stem cells: cell biology and potential use in therapy. , 2004, Basic & clinical pharmacology & toxicology.

[9]  R. Zhao,et al.  Systemic Infusion of FLK1+ Mesenchymal Stem Cells Ameliorate Carbon Tetrachloride-Induced Liver Fibrosis in Mice , 2004, Transplantation.

[10]  M. Grompe,et al.  Myelomonocytic cells are sufficient for therapeutic cell fusion in liver , 2004, Nature Medicine.

[11]  R. Tuan,et al.  Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy , 2004, Journal of cellular and molecular medicine.

[12]  S. Baylin,et al.  Hematopoietic stem cells convert into liver cells within days without fusion , 2004, Nature Cell Biology.

[13]  F. Camargo,et al.  Hematopoietic myelomonocytic cells are the major source of hepatocyte fusion partners. , 2004, The Journal of clinical investigation.

[14]  F. Barry,et al.  Mesenchymal stem cells: clinical applications and biological characterization. , 2004, The international journal of biochemistry & cell biology.

[15]  J. Roy-Chowdhury,et al.  Hepatocyte transplantation: state of the art and strategies for overcoming existing hurdles. , 2004, Annals of hepatology.

[16]  O. Lee,et al.  Isolation of multipotent mesenchymal stem cells from umbilical cord blood. , 2004, Blood.

[17]  S. Forbes,et al.  Hepatic stem cells: from inside and outside the liver? , 2004, Cell proliferation.

[18]  H. Cai,et al.  Expanding hepatocytes in vitro before cell transplantation: donor age‐dependent proliferative capacity of cultured human hepatocytes , 2004, Scandinavian journal of gastroenterology.

[19]  S. Weyers,et al.  Miniature, Low‐Dose, Intrauterine Drug‐Delivery Systems , 2003, Annals of the New York Academy of Sciences.

[20]  Klaus Pfeffer,et al.  Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes , 2003, Nature.

[21]  P. Hayes,et al.  Human cord blood-derived cells can differentiate into hepatocytes in the mouse liver with no evidence of cellular fusion. , 2003, Gastroenterology.

[22]  W. Tan,et al.  Differentiation of putative hepatic stem cells derived from adult rats into mature hepatocytes in the presence of epidermal growth factor and hepatocyte growth factor. , 2003, Differentiation; research in biological diversity.

[23]  J. Wingard,et al.  Transplanted Human Cord Blood Cells Give Rise to Hepatocytes in Engrafted Mice , 2003, Annals of the New York Academy of Sciences.

[24]  G. Vassilopoulos,et al.  Transplanted bone marrow regenerates liver by cell fusion , 2003, Nature.

[25]  M. Grompe,et al.  Cell fusion is the principal source of bone-marrow-derived hepatocytes , 2003, Nature.

[26]  M. Grompe,et al.  Kinetics of liver repopulation after bone marrow transplantation. , 2002, The American journal of pathology.

[27]  R. Schwartz,et al.  Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. , 2002, The Journal of clinical investigation.

[28]  M. Grompe.  Liver repopulation for the treatment of metabolic diseases , 2001, Journal of Inherited Metabolic Disease.

[29]  D. Granner,et al.  Regulation by glucagon (cAMP) and insulin of the promoter of the human phosphoenolpyruvate carboxykinase gene (cytosolic) in cultured rat hepatocytes and in human hepatoblastoma cells. , 2000, The Biochemical journal.

[30]  Xin Wang,et al.  Purified hematopoietic stem cells can differentiate into hepatocytes in vivo , 2000, Nature Medicine.

[31]  D. Melton,et al.  Early mouse endoderm is patterned by soluble factors from adjacent germ layers. , 2000, Development.

[32]  G. Michalopoulos,et al.  Serum-free, long-term cultures of human hepatocytes: maintenance of cell morphology, transcription factors, and liver-specific functions. , 2000, Biochemical and biophysical research communications.

[33]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[34]  S. Strom,et al.  Treatment of the Crigler-Najjar syndrome type I with hepatocyte transplantation. , 1998, The New England journal of medicine.

[35]  Seung Ki Lee,et al.  DL‐propranolol negatively regulates the transcription of proliferating cell nuclear antigen (PCNA)‐gene and thereby suppresses DNA synthesis in regenerating rat liver , 1997, Biochemistry and molecular biology international.

[36]  G. Ramadori,et al.  Cytokines and the hepatic acute phase response , 1997, The Journal of pathology.

[37]  O. Bagasra,et al.  Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[38]  K. Jungermann,et al.  Zonation of metabolism and gene expression in liver , 1995, Histochemistry and Cell Biology.

[39]  O. Bachs,et al.  Possible cyclic AMP‐dependence of the prereplicative surge of cytosolic calmodulin in proliferatively activated rat liver cells , 1988, Journal of cellular physiology.

[40]  L. Blendis,et al.  Reversal of toxic and anoxic induced hepatic failure by syngeneic, allogeneic, and xenogeneic hepatocyte transplantation. , 1980, Surgery.

[41]  D. Wybenga,et al.  Manual and automated methods for urea nitrogen measurement in whole serum. , 1971, Clinical chemistry.

[42]  Catherine M. Verfaillie,et al.  Pluripotency of mesenchymal stem cells derived from adult marrow , 2007, Nature.

[43]  R. Consolini,et al.  Isolation of multipotent mesenchymal stem cells from umbilical cord blood , 2006 .

[44]  A. Zander,et al.  Hepatocytic gene expression in cultured rat mesenchymal stem cells. , 2005, Transplantation proceedings.

[45]  T. Takayama,et al.  Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion , 2005 .

[46]  Jean S. Campbell,et al.  The role of hepatocytes and oval cells in liver regeneration and repopulation , 2003, Mechanisms of Development.

[47]  R. Hanson,et al.  Regulation of phosphoenolpyruvate carboxykinase (GTP) gene expression. , 1997, Annual review of biochemistry.