title: Enhanced proliferation and differentiation of rat hepatocytes cultured with bone marrow stromal cells.
abstract: Liver transplantation is the only clinically effective method of treating acute liver failure. However, wider application of this therapeutic modality is restricted primarily by shortage of donor organs. In the search for alternative methods of liver replacement therapy, investigators have focused on transplantation of normal allogeneic hepatocytes and on the development of liver support systems utilizing isolated hepatocytes. Since all human livers suitable for cell harvest are being used for transplantation, hepatocyte therapy using human tissue would require growing of cells in vitro. Unfortunately, although hepatocytes have tremendous capacity to proliferate in vivo, their ability to grow in culture is severely limited. Stromal cells from bone marrow and other blood-forming organs have been found to support hematopoiesis. In this paper, we show that bone marrow-derived stromal cells (BMSCs) enhance proliferation and support differentiation of rat hepatocytes in culture. Further, we demonstrate that in hepatocyte/BMSC co-cultures, clonal expansion of small hepatocytes (SH) is increased. Using semipermeable membrane cultures, we established that direct cell-cell contact is necessary for stimulation of cell proliferation. We also show that BMSCs which are in direct contact with hepatocytes and SH colonies express Jagged1. This suggests a potential role for Notch signaling in the observed effects. Finally, we present evidence that the expression and activity of liver specific transcription factors, CCAAT/enhancer binding proteins and liver specific key enzymes such as tryptophan 2,3-dioxygenase, are improved in hepatocyte/BMSC co-cultures. In conclusion, results of this study indicate that BMSCs could facilitate proliferation and differentiation of primary rat hepatocytes and their progenitors (SH) in vitro.
authors.affiliation: Liver Support Research Laboratory, Cedars-Sinai Medical Center, UCLA School of Medicine, Los Angeles, California 90048, USA. mizuguchit@cshs.org
authors.forename: T
authors.initials: T
authors.lastname: Mizuguchi
authors.forename: T
authors.initials: T
authors.lastname: Hui
authors.forename: K
authors.initials: K
authors.lastname: Palm
authors.forename: N
authors.initials: N
authors.lastname: Sugiyama
authors.forename: T
authors.initials: T
authors.lastname: Mitaka
authors.forename: A A
authors.initials: AA
authors.lastname: Demetriou
authors.forename: J
authors.initials: J
authors.lastname: Rozga
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D001973:Bromodeoxyuridine / Q000378:metabolism; D022762:CCAAT-Enhancer-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D002135:Calcium-Binding Proteins; D002450:Cell Communication; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D000072100:Jagged-1 Protein; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins; D010641:Phenotype; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D000072099:Serrate-Jagged Proteins; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D017154:Stromal Cells / Q000502:physiology*
chemical_list: D022762:CCAAT-Enhancer-Binding Proteins; D002135:Calcium-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; C000606275:Jag1 protein, rat; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D011506:Proteins; D012333:RNA, Messenger; D000072099:Serrate-Jagged Proteins; D001973:Bromodeoxyuridine