| ObjectiveWithin the bone marrow stroma there exists a subset of nonhematopoietic stem cells referred to as marrow stromal cells or mesenchymal stem cells. Mesenchymal stem cells (MSCs) are a group of cells with highly capability of self-renew and potential of multilineage differentiation, these properties make them present a promising prospect for clinical practice. Here we observe the biological features of MSCs , explore whether adult mesenchymal stem cells can be induced to develop into hepatocytes and the role of hepatocyte growth factor(HGF) combined with basic fibroblast factor (bFGF) and fetal liver cells in inducing MSCs differentiating into hepatocytes in vitro. The purpose of this study was to provide new source of cells for liver cell transplantation and bioartificial liver therapy. Methods1. The bone marrow was aspirated from the iliac crest of healthy donors and MSCs was isolated by density gradient centrifugation in percoll. The mononuclear cells were collected and cultured in vitro, and the adherent cells were kept. The higher purity of MSCs was obtained by repeatedly removal of nonadherent cells. The biological features were observed under phase contrast microscope.2. Liver obtained from legally aborted fetuses between 2—4 month of gestation was digested with 0.05% collagenase I and the higher purity of liver cells were obtained by changing the media after 4h initial inoculation . The biological features were observed under phase contrast microscope.3. To induce hepatic differentiation effectively, MSCs between passages 4 and 8 were induced with the use of hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF) . The morphological changes of induced MSCs were observed in different culture period. The hepatic surface phenotype including AFP, CK18 and CK19 were identified by immunofluorescent staining at day7, 14, 21, 28. Glycogen storage was detected by Periodic Acid-Schiff(PAS) staining and the... |
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