Conditional TGF-β1 treatment increases stem cell-like cell population in myoblasts.
Bottom Line: The limitation in successfully acquiring large populations of stem cell has impeded their application.In addition to the C2C12 myoblasts, similar effects of TGF-β(1) were also observed in the primary myoblasts of mice.Our results suggest that TGF-β(1) is effective as a molecular trigger for the dedifferentiation of skeletal muscle myoblasts and could be used to generate a large pool of progenitor cells that collectively behave as multipotent stem cell-like cells for regenerative medicine applications.
Affiliation: The Laboratory of Molecular Pathology, Stem Cell Research Center, Children's Hospital of UPMC, Pittsburgh, PA, USA.Show MeSH
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Mentions: We also investigated the osteogenic potential of the dedifferentiated cells, in order to confirm that the cells can differentiate into other cell lineages. hrTGF-β1 pre-treated (0.5 ng/ml, 3 hrs) and non-treated C2C12 myoblasts were cultured in osteogenic medium for 4 days before being fixed for detecting the activity of ALP, an early marker of osteogenesis . Results showed that the TGF-β1 pre-treated cells demonstrated a higher ALP signal, indicating a stronger osteogenic potential of dedifferentiated cells compared to the control cells (Fig. 5A, B and E) in vitro. In vivo, we created a 6-mm-diameter defect in the parietal bone of SCID mice without breaching the dura, and then a 7 mm Gelfoam disk impregnated with either 2 × 105 hrTGF-β1 pre-treated (0.5 ng/ml, 3 hrs) or non-treated C2C12 myoblasts was transplanted into the defect. Bone healing was monitored by microCT (vivaCT40, Scanco Medical AG). Small bone tissues were observed in the defect area as shown by micro CT at 8 weeks after surgery (arrows, Fig. 5C and D), and the TGF-β1 pre-treated C2C12 obviously formed more bone tissue than the control cells (Fig. 5C, D and F).
Affiliation: The Laboratory of Molecular Pathology, Stem Cell Research Center, Children's Hospital of UPMC, Pittsburgh, PA, USA.