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Clones of ectopic stem cells in the regeneration of muscle defects in vivo.

Yang R, Chen M, Lee CH, Yoon R, Lal S, Mao JJ - PLoS ONE (2010)

Bottom Line: The fusion indices of myogenic clones were approximately 16-17 folds greater than their parent, heterogeneous stem cells.Strikingly, clonal progenies yielded ∼9 times more human myosin heavy chain mRNA in regenerating muscles than those infused with their parent, heterogeneous stem cells.The number of human dystrophin positive cells in regenerating muscles infused with clonal progenies was more than ∼3 times greater than muscles infused with heterogeneous stem cells from which clonal progenies were derived.

View Article: PubMed Central - PubMed

Affiliation: Tissue Engineering and Regenerative Medicine Laboratory, Department of Growth and Development, Columbia University Medical Center, New York, New York, United States of America.

ABSTRACT
Little is known about whether clones of ectopic, non-muscle stem cells contribute to muscle regeneration. Stem/progenitor cells that are isolated for experimental research or therapeutics are typically heterogeneous. Non-myogenic lineages in a heterogeneous population conceptually may compromise tissue repair. In this study, we discovered that clones of mononucleated stem cells of human tooth pulp fused into multinucleated myotubes that robustly expressed myosin heavy chain in vitro with or without co-culture with mouse skeletal myoblasts (C2C12 cells). Cloned cells were sustainably Oct4+, Nanog+ and Stro1+. The fusion indices of myogenic clones were approximately 16-17 folds greater than their parent, heterogeneous stem cells. Upon infusion into cardio-toxin induced tibialis anterior muscle defects, undifferentiated clonal progenies not only engrafted and colonized host muscle, but also expressed human dystrophin and myosin heavy chain more efficaciously than their parent heterogeneous stem cell populations. Strikingly, clonal progenies yielded ∼9 times more human myosin heavy chain mRNA in regenerating muscles than those infused with their parent, heterogeneous stem cells. The number of human dystrophin positive cells in regenerating muscles infused with clonal progenies was more than ∼3 times greater than muscles infused with heterogeneous stem cells from which clonal progenies were derived. These findings suggest the therapeutic potential of ectopic myogenic clones in muscle regeneration.

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Myogenic potential of clonal progeny of ectopic dental stem cells in chemically defined medium.A: Phase contrast image of a representative heterogeneous dental stem cell (DSC) population showing typical spindle-shaped cells. Upon exposure to chemically defined medium, DSCs assumed spherical shape but failed to fuse into multinucleated cells (B) or express myosin heavy chain (C). Strikingly, two representative clones (B6 and C3) in chemically defined medium for 4 wks readily fused into tubular structures (D,E) that are positive for myosin heavy chain (MHC) immunoblotting (F,G). Quantitatively, MHC mRNA expression is significantly greater by B6 and C3 cells than their heterogeneous parent cells (DSC) (H). The y axis represents fold change related to heterogeneous DSC. Scale bar: 100 µm.
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pone-0013547-g003: Myogenic potential of clonal progeny of ectopic dental stem cells in chemically defined medium.A: Phase contrast image of a representative heterogeneous dental stem cell (DSC) population showing typical spindle-shaped cells. Upon exposure to chemically defined medium, DSCs assumed spherical shape but failed to fuse into multinucleated cells (B) or express myosin heavy chain (C). Strikingly, two representative clones (B6 and C3) in chemically defined medium for 4 wks readily fused into tubular structures (D,E) that are positive for myosin heavy chain (MHC) immunoblotting (F,G). Quantitatively, MHC mRNA expression is significantly greater by B6 and C3 cells than their heterogeneous parent cells (DSC) (H). The y axis represents fold change related to heterogeneous DSC. Scale bar: 100 µm.

Mentions: In contrast to their typical spindle shape (Fig. 3A), mononucleated and adherent cells of the tooth pulp upon 4-wk exposure to chemically defined myogenic differentiation medium assumed a somewhat rounded morphology (Fig. 3B). However, these heterogeneous stem/progenitor cells failed to fuse into multinucleated cells or express MHC (Fig. 3C). Strikingly, B6 and C3 exposed for 4 wks in the same chemically defined medium readily fused into multi-nucleated myotube-like structures (Fig. 3D,E) and robustly expressed MHC (Fig. 3F,G). Remarkably, fusion into multi-nucleated, MHC+ cells occurred in 4 wks without co-culture with mouse skeletal myoblasts (C2C12). Overall, ∼27% of B6 and ∼30% of C3 cells fused into multi-nucleated, MHC+ myotube-like structures that were quantified from multiple plates. Quantitative real-time PCR analysis revealed significantly greater MHC expression by B6 and C3 cells heterogeneous DSCs, from which B6 and C3 were cloned (Fig. 3H).


Clones of ectopic stem cells in the regeneration of muscle defects in vivo.

Yang R, Chen M, Lee CH, Yoon R, Lal S, Mao JJ - PLoS ONE (2010)

Myogenic potential of clonal progeny of ectopic dental stem cells in chemically defined medium.A: Phase contrast image of a representative heterogeneous dental stem cell (DSC) population showing typical spindle-shaped cells. Upon exposure to chemically defined medium, DSCs assumed spherical shape but failed to fuse into multinucleated cells (B) or express myosin heavy chain (C). Strikingly, two representative clones (B6 and C3) in chemically defined medium for 4 wks readily fused into tubular structures (D,E) that are positive for myosin heavy chain (MHC) immunoblotting (F,G). Quantitatively, MHC mRNA expression is significantly greater by B6 and C3 cells than their heterogeneous parent cells (DSC) (H). The y axis represents fold change related to heterogeneous DSC. Scale bar: 100 µm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2958121&req=5

pone-0013547-g003: Myogenic potential of clonal progeny of ectopic dental stem cells in chemically defined medium.A: Phase contrast image of a representative heterogeneous dental stem cell (DSC) population showing typical spindle-shaped cells. Upon exposure to chemically defined medium, DSCs assumed spherical shape but failed to fuse into multinucleated cells (B) or express myosin heavy chain (C). Strikingly, two representative clones (B6 and C3) in chemically defined medium for 4 wks readily fused into tubular structures (D,E) that are positive for myosin heavy chain (MHC) immunoblotting (F,G). Quantitatively, MHC mRNA expression is significantly greater by B6 and C3 cells than their heterogeneous parent cells (DSC) (H). The y axis represents fold change related to heterogeneous DSC. Scale bar: 100 µm.
Mentions: In contrast to their typical spindle shape (Fig. 3A), mononucleated and adherent cells of the tooth pulp upon 4-wk exposure to chemically defined myogenic differentiation medium assumed a somewhat rounded morphology (Fig. 3B). However, these heterogeneous stem/progenitor cells failed to fuse into multinucleated cells or express MHC (Fig. 3C). Strikingly, B6 and C3 exposed for 4 wks in the same chemically defined medium readily fused into multi-nucleated myotube-like structures (Fig. 3D,E) and robustly expressed MHC (Fig. 3F,G). Remarkably, fusion into multi-nucleated, MHC+ cells occurred in 4 wks without co-culture with mouse skeletal myoblasts (C2C12). Overall, ∼27% of B6 and ∼30% of C3 cells fused into multi-nucleated, MHC+ myotube-like structures that were quantified from multiple plates. Quantitative real-time PCR analysis revealed significantly greater MHC expression by B6 and C3 cells heterogeneous DSCs, from which B6 and C3 were cloned (Fig. 3H).

Bottom Line: The fusion indices of myogenic clones were approximately 16-17 folds greater than their parent, heterogeneous stem cells.Strikingly, clonal progenies yielded ∼9 times more human myosin heavy chain mRNA in regenerating muscles than those infused with their parent, heterogeneous stem cells.The number of human dystrophin positive cells in regenerating muscles infused with clonal progenies was more than ∼3 times greater than muscles infused with heterogeneous stem cells from which clonal progenies were derived.

View Article: PubMed Central - PubMed

Affiliation: Tissue Engineering and Regenerative Medicine Laboratory, Department of Growth and Development, Columbia University Medical Center, New York, New York, United States of America.

ABSTRACT
Little is known about whether clones of ectopic, non-muscle stem cells contribute to muscle regeneration. Stem/progenitor cells that are isolated for experimental research or therapeutics are typically heterogeneous. Non-myogenic lineages in a heterogeneous population conceptually may compromise tissue repair. In this study, we discovered that clones of mononucleated stem cells of human tooth pulp fused into multinucleated myotubes that robustly expressed myosin heavy chain in vitro with or without co-culture with mouse skeletal myoblasts (C2C12 cells). Cloned cells were sustainably Oct4+, Nanog+ and Stro1+. The fusion indices of myogenic clones were approximately 16-17 folds greater than their parent, heterogeneous stem cells. Upon infusion into cardio-toxin induced tibialis anterior muscle defects, undifferentiated clonal progenies not only engrafted and colonized host muscle, but also expressed human dystrophin and myosin heavy chain more efficaciously than their parent heterogeneous stem cell populations. Strikingly, clonal progenies yielded ∼9 times more human myosin heavy chain mRNA in regenerating muscles than those infused with their parent, heterogeneous stem cells. The number of human dystrophin positive cells in regenerating muscles infused with clonal progenies was more than ∼3 times greater than muscles infused with heterogeneous stem cells from which clonal progenies were derived. These findings suggest the therapeutic potential of ectopic myogenic clones in muscle regeneration.

Show MeSH
Related in: MedlinePlus