ABSTRACT
BACKGROUND AIMS: Mesenchymal stromal cells (MSC) isolated from bone marrow (BM), adipose tissue and umbilical cord blood can be induced to differentiate into hepatocyte-like cells. MSC can also be isolated from umbilical cord Wharton's jelly (UC MSC), which can be easily obtained. UC MSC are more primitive MSC than those isolated from other tissue sources and do not express the major histocompatibility complex (MHC) class II (HLA-DR) antigens. Previous studies have shown that UC MSC are still viable and not rejected 4 months after transplantation as xenografts, without the need for immune suppression, suggesting that they are a favorable cell source for transplantation. METHODS: UC MSC were induced to differentiate into hepatocyte-like cells by a simple one-step protocol with hepatotic growth factor (HGF) and fibroblast growth factor-4 (FGF-4). Differentiated cells were examined for the expression of hepatocyte-specific markers and hepatocyte functions. RESULTS: UC MSC were isolated. Flow cytometry analysis showed that they expressed the MSC-specific markers. They differentiated into osteoblast-, adipocyte- and chondrocyte-like cells, showing their multipotent differentiation potential. Immunocytochemistry, real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis demonstrated that UC MSC expressed the hepatocyte-specific markers albumin (ALB), human alpha-fetoprotein (AFP) and cytokeratin 18 (CK-18) following hepatocyte induction. Periodic acid-Schiff staining showed that differentiated UC MSC could store glycogen, and an low-density lipoprotein (LDL)-uptake assay showed that they could uptake LDL. CONCLUSIONS: This study demonstrates that UC MSC can differentiate into functional hepatocyte-like cells following the induction of HGF and FGF-4. UC MSC can serve as a favorable cell source for tissue engineering in the treatment of liver disease.
Subject(s)
Cell Differentiation , Hepatocytes/cytology , Mesenchymal Stem Cells/cytology , Stromal Cells/cytology , Umbilical Cord/cytology , Adipogenesis , Biomarkers/metabolism , Blotting, Western , Cell Separation , Cells, Cultured , Chondrogenesis , Female , Flow Cytometry , Gene Expression Regulation, Developmental , Hepatocytes/metabolism , Humans , Immunohistochemistry , Lipoproteins, LDL/metabolism , Osteogenesis , Periodic Acid-Schiff Reaction , Pregnancy , Reverse Transcriptase Polymerase Chain Reaction , Staining and Labeling , Time FactorsABSTRACT
OBJECTIVE: Mesenchymal stem cells (MSCs) can be isolated from umbilical cord Wharton's jelly (UC-MSC) and UC can be easily obtained, representing a noncontroversial source of MSCs. UC-MSCs are more primitive than other tissue sources. Previous studies showed that UC-MSCs were still viable and were not rejected 4 months after transplantation as xenografts without the need for immune suppression, indicating that they are favorable cell source for transplantation. In this study, UC-MSCs were induced to differentiate into endothelial-like cells and compared with bone marrow (BM)-MSCs for their endothelial differentiation potential. MATERIALS AND METHODS: UC-MSCs and BM-MSCs were characterized for expression of MSC-specific markers and osteogenic, adipogenic, and chondrogenic differentiation. They were induced to differentiate into endothelial-like cells and analyzed for expression of the endothelial-specific markers and functions. RESULTS: UC-MSCs and BM-MSCs showed similarities in expression of the MSC-specific markers and osteogenic, adipogenic, and chondrogenic differentiation. They showed similar low-density lipoprotein-uptaking capacity following endothelial differentiation. However, UC-MSCs had higher proliferative potential than BM-MSCs. Both real-time reverse transcription polymerase chain reaction and immunocytochemical analyses demonstrated that UC-MSCs had higher expression of the endothelial-specific markers than BM-MSCs following endothelial differentiation. Both Matrigel and coculture angiogenesis assays showed that UC-MSCs and BM-MSCs after endothelial differentiation were able to form the capillary network and differentiated UC-MSCs had significantly higher total tubule length, diameter, and area than differentiated BM-MSCs. CONCLUSION: These results showed that UC-MSCs had higher endothelial differentiation potential than BM-MSCs. Therefore, UC-MSCs are more favorable choice than BM-MSCs for neovascularization of engineered tissues.
