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Conditional TGF-β1 treatment increases stem cell-like cell population in myoblasts.

Mu X, Li Y - J. Cell. Mol. Med. (2011)

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.

View Article: PubMed Central - PubMed

Affiliation: The Laboratory of Molecular Pathology, Stem Cell Research Center, Children's Hospital of UPMC, Pittsburgh, PA, USA.

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Osteogenic potential of TGF-β1 pre-treated C2C12 myoblasts in vitro and their efficiency in repair skull defect in vivo. In vitro osteogenic differentiation was compared between C2C12 control cells (A) and cells pre-treated with hrTGF-β1 (0.5 ng/ml) (B), by measuring the percentage of cells positive with ALP signal. The efficiency of bone tissue formation in vivo was compared between C2C12 control cells (C) and cells pre-treated with TGF-β1 (D). Four mice in each group (control cells and TGF-β1 treated cells) were included in this study. Some small bone tissue was observed in the defect area at 8 weeks after surgery by micro CT, and TGF-β1 pre-treated C2C12 myoblasts demonstrated a higher efficiency in repairing injured bone (D) compared to the control cells (C). Statistical quantitation of ALP+ cells in vitro (E) and bone tissue formation in vivo was included (F). ‘*’ indicates the value being significantly different with the control cells (TGF-β1–).
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fig05: Osteogenic potential of TGF-β1 pre-treated C2C12 myoblasts in vitro and their efficiency in repair skull defect in vivo. In vitro osteogenic differentiation was compared between C2C12 control cells (A) and cells pre-treated with hrTGF-β1 (0.5 ng/ml) (B), by measuring the percentage of cells positive with ALP signal. The efficiency of bone tissue formation in vivo was compared between C2C12 control cells (C) and cells pre-treated with TGF-β1 (D). Four mice in each group (control cells and TGF-β1 treated cells) were included in this study. Some small bone tissue was observed in the defect area at 8 weeks after surgery by micro CT, and TGF-β1 pre-treated C2C12 myoblasts demonstrated a higher efficiency in repairing injured bone (D) compared to the control cells (C). Statistical quantitation of ALP+ cells in vitro (E) and bone tissue formation in vivo was included (F). ‘*’ indicates the value being significantly different with the control cells (TGF-β1–).

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 [54]. 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).


Conditional TGF-β1 treatment increases stem cell-like cell population in myoblasts.

Mu X, Li Y - J. Cell. Mol. Med. (2011)

Osteogenic potential of TGF-β1 pre-treated C2C12 myoblasts in vitro and their efficiency in repair skull defect in vivo. In vitro osteogenic differentiation was compared between C2C12 control cells (A) and cells pre-treated with hrTGF-β1 (0.5 ng/ml) (B), by measuring the percentage of cells positive with ALP signal. The efficiency of bone tissue formation in vivo was compared between C2C12 control cells (C) and cells pre-treated with TGF-β1 (D). Four mice in each group (control cells and TGF-β1 treated cells) were included in this study. Some small bone tissue was observed in the defect area at 8 weeks after surgery by micro CT, and TGF-β1 pre-treated C2C12 myoblasts demonstrated a higher efficiency in repairing injured bone (D) compared to the control cells (C). Statistical quantitation of ALP+ cells in vitro (E) and bone tissue formation in vivo was included (F). ‘*’ indicates the value being significantly different with the control cells (TGF-β1–).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3008543&req=5

fig05: Osteogenic potential of TGF-β1 pre-treated C2C12 myoblasts in vitro and their efficiency in repair skull defect in vivo. In vitro osteogenic differentiation was compared between C2C12 control cells (A) and cells pre-treated with hrTGF-β1 (0.5 ng/ml) (B), by measuring the percentage of cells positive with ALP signal. The efficiency of bone tissue formation in vivo was compared between C2C12 control cells (C) and cells pre-treated with TGF-β1 (D). Four mice in each group (control cells and TGF-β1 treated cells) were included in this study. Some small bone tissue was observed in the defect area at 8 weeks after surgery by micro CT, and TGF-β1 pre-treated C2C12 myoblasts demonstrated a higher efficiency in repairing injured bone (D) compared to the control cells (C). Statistical quantitation of ALP+ cells in vitro (E) and bone tissue formation in vivo was included (F). ‘*’ indicates the value being significantly different with the control cells (TGF-β1–).
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 [54]. 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).

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.

View Article: PubMed Central - PubMed

Affiliation: The Laboratory of Molecular Pathology, Stem Cell Research Center, Children's Hospital of UPMC, Pittsburgh, PA, USA.

Show MeSH
Related in: MedlinePlus