Nuclear Translocation of β-Catenin during Mesenchymal Stem Cells Differentiation into Hepatocytes Is Associated with a Tumoral Phenotype
Figure 5
A) Number of cells after 7, 14 and 21 days of culture in undifferentiated, CM1 and CM2-treated cells. Values expressed as mean ± standard deviation. a p<0.001 vs. CM1-treated cells and undifferentiated cells. B) The presence of nuclear PCNA (brown nucleus) was evaluated by immunohistochemistry after 21 days of culture in undifferentiated cells, CM and CM2-treated cells. Image is representative of three experiments. Original magnification: 20×. C) Cell cycle was analyzed at 21 days of hepatocyte differentiation in undifferentiated, CM1 and CM2-tretaed cells. Data are showed as mean of percentage plus standard deviation. a p<0.001 vs. CM1-treated cells, ++ p<0.01 and +++ p<0.001 vs. undifferentiated cells. D) Primary spheroid assay with count of number of cells after 4 days of culture with conditioned medium for spheroid formation. Data are showed that mean ± standard deviation (a p<0.001 vs. CM1-treated cells and +++ p<0.001 vs. undifferentiated cells). E) Secondary spheroid formation assay. Number of secondary spheroids was counted in an inverted microscope. Three experiments were carried out and data are expressed as mean ± standard deviation (a p<0.001 vs. CM1-treated cells and +++ p<0.001 vs. undifferentiated cells). F) Detail of secondary spheroids is showed in the microphotographs of undifferentiated cells, CM1 and CM2-treated cells.