Supplementary Figure 7: Characterization of MSCs and MSC-derived cells.

(a) FACS results of the affected patient VI/17 and one non-affected control. The analysis of surface markers detected CD105⁺, CD90⁺, CD73⁺, HLA-ABC⁺, CD31^–, CD34^–, CD45^– and HLA-DR^– cells. (b) Plastic adherence and the multilineage potential of MSCs of one non-affected control and VI/17. MSCs and VSMCs from patient VI/9 and the second non-affected control were identically characterized and fulfilled all criteria of MSCs and VSMCs (data not shown). Immunocytochemical staining of the MSC-derived adipocytes, osteocytes and chondrocytes validated the multilineage potential of the MSCs. The fatty vacuoles show successful differentiation into adipocytes; calcium precipitates characterized MSC-derived osteocytes. Toluidine blue stained proteoglycans of sectioned chondrogenic tissue that was generated in micromass pellet cultures. Successful differentiation was similar in the MSCs from the control and affected patient VI/17. MSC characterizations fulfilled the criteria of the International Society for Cellular Therapy. (c) Myogenic differentiation of one non-affected control and MSCs from VI/17 into VSMCs after 21 d of differentiation. Semiquantitative immunofluorescence detected the smooth muscle markers smooth muscle actin (SMAα), calponin and transgelin (SM22α), which were highly expressed compared to in undifferentiated MSCs. The color spectrum indicates the expression level from low (black) to high (red).