title: Fetal liver-derived mesenchymal stromal cells augment engraftment of transplanted hepatocytes.
abstract: OBJECTIVE
  One important problem commonly encountered after hepatocyte transplantation is the low numbers of transplanted cells found in the graft. If hepatocyte transplantation is to be a viable therapeutic approach, significant liver parenchyma repopulation is required. Mesenchymal stromal cells (MSC) produce high levels of various growth factors, cytokines and metalloproteinases, and have immunomodulatory effects. We therefore hypothesized that co-transplantation of MSC with human fetal hepatocytes (hFH) could augment in vivo expansion after transplantation. We investigated the ability of human fetal liver MSC (hFLMSC) to augment expansion of phenotypically and functionally well-characterized hFH.
  
  
  METHODS
  Two million hFH (passage 6) were either transplanted alone or together (1:1 ratio) with green fluorescence protein-expressing hFLMSC into the spleen of C57BL/6 nude mice with retrorsine-induced liver injury.
  
  
  RESULTS
  After 4 weeks, engraftment of cells was detected by fluorescence in situ hybridization using a human-specific DNA probe. Significantly higher numbers of cells expressing human cytokeratin (CK)8, CK18, CK19, Cysteine-rich MNNG HOS Transforming gene (c-Met), alpha-fetoprotein (AFP), human nuclear antigen, mitochondrial antigen, hepatocyte-specific antigen and albumin (ALB) were present in the livers of recipient animals co-transplanted with hFLMSC compared with those without. Furthermore, expression of human hepatocyte nuclear factor (HNF)-4α and HNF-1β, and cytochrome P450 (CYP) 3A7 mRNA was demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR) in these animals. In addition, significantly increased amounts of human ALB were detected. Importantly, hFLMSC did not transdifferentiate into hepatocytes.
  
  
  CONCLUSIONS
  Our study reports the use of a novel strategy for enhanced liver repopulation and thereby advances this experimental procedure closer to clinical liver cell therapy.
authors.affiliation: Department of Surgery, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden.
authors.forename: Meghnad
authors.initials: M
authors.lastname: Joshi
authors.forename: Pradeep
authors.initials: P
authors.lastname: B Patil
authors.forename: Zhong
authors.initials: Z
authors.lastname: He
authors.forename: Jan
authors.initials: J
authors.lastname: Holgersson
authors.forename: Michael
authors.initials: M
authors.lastname: Olausson
authors.forename: Suchitra
authors.initials: S
authors.lastname: Sumitran-Holgersson
mesh_terms: D000818:Animals; D000941:Antigens / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism; D005333:Fetus / Q000166:cytology*; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation*; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000196:embryology*; D008107:Liver Diseases / Q000473:pathology / Q000628:therapy; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D010641:Phenotype; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D011763:Pyrrolizidine Alkaloids; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012709:Serum Albumin / Q000378:metabolism; D013045:Species Specificity; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D000941:Antigens; D015415:Biomarkers; D018925:Chemokines; D011763:Pyrrolizidine Alkaloids; D012333:RNA, Messenger; D012709:Serum Albumin; D019859:Proto-Oncogene Proteins c-met; C003300:retrorsine