Objective: To explore the ability of QY1 bone marrow mesenchymal stem cell (MSCs) line cells to differentiate into adipocytes, chondrocytes, osteoblasts, cardiac myocytes,vascular endothelial cells, and neural cells in vitro.
Methods: The QY1 cells at passage 5 were treated with the adipogenic medium, the chondrogenic medium and the osteogenic medium, 5-azacytidine, vascular endothelial growth factor and neural cell medium (revulsant 1 was 10 mmol/L beta-mercaptoethanol; revulsant 2 was 2%dimethylsulfoxide and 10(-8)mol/L dexamethasone) in culture respectively in vitro. The differentiated cells were identified by staining, immunohistochemistry and RT-PCR.
Results: The differentiated cells induced by the adipogenic medium formed adipocytes and contained fat lipid droplets, which were stained positively with Sudan III after 21 days of culture. The differentiated cells induced by the chondrogenic medium formed chondrogenic nodules, which were stained positively by Alcian blue at pH 1.0 after 21 days of culture. The differentiated cells induced by the osteogenic medium formed osteogenic nodules, which were stained positively by Von Kossa staining after 35 days of culture, and the secretion of a calcified extracellular matrix as black nodules was observed. The differentiated cells treated with 10 micromol/L 5-azacytidine could beat spontaneously and formed myotube structures,which were identified by the positive immunohistochemistry staining with anti-alpha-sarcomeric antibody and anti-Cx-43 antibody. The expression of alpha-myosin heavy chain was also observed by RT-PCR. The differentiated cells treated with 50 ng/mL vascular endothelial growth factor could form vascular endothelial cells and vascular endothelial web like structure, which were identified by the positive immunohistochemistry staining with CD31 and Factor VIII. The differentiated cells induced by revulsant 1 were positive in the immunohistochemistry staining with neuron-specific nuclear protein, while the expression of glial fibrillary acidic protein was negative. The differentiated cells induced by revulsant 2 were positive in the immunohistochemistry staining with glial fibrillary acidic protein, while the expression of neuron-specific nuclear protein was negative.
Conclusion: QY1 bone marrow mesenchymal stem cell line has the ability to differentiate into adipocytes, chondrocytes, osteocytes, cardiomyocytes, vascular endothelial cells, neurons and neural glial cells in vitro. A bone marrow mesenchymal stem cell line cell can at least differentiate into 7 types of cells, which come from mesoderm and ectoderm.