Persistence of a chimerical phenotype after hepatocyte differentiation of human bone marrow mesenchymal stem cells.
OBJECTIVE
Recent studies have suggested the potential of mesenchymal stem cells (MSCs) to differentiate into a hepatocyte-like lineage. Here, we evaluate the efficacy of hepatocyte differentiation of MSCs by studying acquisition of hepatocyte-like features together with alteration of the native mesenchymal phenotype.


METHODS
In vitro, we have investigated protein and mRNA level expression of hepatocyte and mesenchymal markers of mesenchymal-derived hepatocyte-like cells (MDHLCs) and we have evaluated their functionality using metabolic assays. In vivo, we investigated co-expression of hepatocyte (albumin, alpha-foetoprotein, cytokeratin 18) and mesenchymal (fibronectin, vimentin) markers after transplantation of MSCs or MDHLCs into severe combined immune deficiency mice.


RESULTS
We observed that while in vitro these cells acquired some phenotypic and functional features of mature hepatocytes, they partially preserved their mesenchymal phenotype. After intrasplenic transplantation, engrafted MSCs with isolated expression of fibronectin and alpha-foetoprotein were observed. When these cells were injected into the liver, they expressed all analysed markers, confirming the chimaeric co-expression observed in vitro. Conversely, liver-engrafted MDHLCs conserved their hepatocyte-lineage markers but lost their chimaeric phenotype.


CONCLUSIONS
Hepatocyte differentiation of MSCs predominantly allows the acquisition of phenotypic hallmarks and provides chimaeric cells that maintain expression of initial lineage markers. However, advanced maturation to the hepatocyte-like phenotype could be obtained in vivo by conditioning MSCs prior to transplantation or by infusing cells into the liver micro-environment.
Lysy
P A
PA
Université Catholique de Louvain & Cliniques Universitaires Saint Luc, HPED Department, PEDI Unit, Laboratory of Pediatric Hepatology and Cell Therapy, Brussels, Belgium. philippe.lysy@pedi.ucl.ac.be
Campard
D
D
Smets
F
F
Malaise
J
J
Mourad
M
M
Najimi
M
M
Sokal
E M
EM
D001483:Base Sequence; D001854:Bone Marrow Cells / Q000166:cytology* / Q000648:ultrastructure; D002454:Cell Differentiation; D002478:Cells, Cultured; D017931:DNA Primers; D005434:Flow Cytometry; D022781:Hepatocytes / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers