Figure 1
Characterization of liver-derived mesenchymal stromal cells by phenotype and expression and localization of Notch receptors and ligands in undifferentiated MSCs and during their differentiation to osteoblasts. (a) Detection of cell surface markers (CD34 and CXCR4) specific for pluripotent rodent MSCs in undifferentiated liver-derived MSCs by flow cytometry. (b) Detection of mRNAs encoding the four Notch receptors and their three ligands in the undifferentiated liver-derived MSCs. Lanes: M—DNA ladder; Notch 1, 2, 3, 4—products reflecting Notch1, 2, 3, and 4, respectively, Ligand, D1, J1, J2—products reflecting DLL1, Jagged1 and 2, respectively. The numbers under the bands indicate the expected length of the PCR product. The question mark denotes an unspecific product obtained for Jagged2. (c) Immunodetection and location of receptors: Notch1, 2, and 3 and ligand Dll1 in liver-derived MSCs differentiating to osteoblasts. The lowest panels show representative microphotographs of specimens with cells cultured under osteoblastic differentiation conditions stained with Alizarin Red S to determine the content of calcium phosphate (Ca2+). N.A.—indicates that Alizarin Red S staining was not performed for this time point. Arrows—cells with the detected antigen in the nucleus or in both the nucleus and cytoplasm. Arrowheads—cells with the detected antigen exclusively in the cytoplasm. Scale bars indicate 100 μm. MSCs, mesenchymal stromal cells.
