Abstract
Purpose
To investigate the production of bile duct endothelial cells via directed differentiation of porcine bone marrow mesenchymal stem cells (BMSCs) down the hepatic lineage in vitro and the biocompatibility of differentiated bile duct endothelial cells with electrospun nanofibers.
Methods
Porcine BMSCs were differentiated in vitro into bile duct endothelial cells, which were identified by morphology and RT-PCR. PLGA nanofiber membranes were prepared by electrospinning. The morphology was detected by scanning electron microscopy and the short-term (two weeks) in vitro degradation rate was determined. Adhesion and proliferation of the bile duct endothelial cells on the nanofiber surface were analyzed by calculating the cell adhesion rate and MTT assay, respectively. Cell growth, morphology and distribution on the material surface were observed by fluorescence staining and scanning electron microscopy, respectively.
Results
After four weeks of directed differentiation of BMSCs in vitro, cells showed the typical morphology of dendritic bile duct endothelial cells and had the expression of CK19. Scanning electron micrographs showed that electrospun materials were continuous nanofibers with diameters between 200 and 500 nm. No significant degradation of the PLGA nanofibers was observed within two weeks. Based on the measured cell adhesion rate, MTT assay, fluorescence staining, and scanning electron microscopy, the differentiated cells possess a good proliferative capacity on PLGA nanofibers.
Conclusions
BMSCs can be differentiated into the bile duct endothelial cells in vitro. Materials prepared by the electrospinning method have a nanofiber structure, which does not significantly degrade within two weeks. Differentiated cells exhibit good biocompatibility with the nanofibers.
Keywords
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