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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 upon co-culture with mouse skeletal myoblasts.Upon co-culture with mouse myoblast cell line (C2C12 cells) for 1 wk, some of the B6 and C3 cells again fused into multinucleated cells (A,B). Human nuclear staining indicates that the presence of human nuclei in some of the fused myotube like, MHC positive structures (A,B). Quantitatively, human nuclear fusion index (C) and human MHC mRNA expression of B6 and C3 (D) are significantly greater than DSC+C2C12 group. In (D), y axis represents fold change related to DSC+C2C12 group. Scale bar, 50 µm.
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pone-0013547-g004: Myogenic potential of clonal progeny of ectopic dental stem cells upon co-culture with mouse skeletal myoblasts.Upon co-culture with mouse myoblast cell line (C2C12 cells) for 1 wk, some of the B6 and C3 cells again fused into multinucleated cells (A,B). Human nuclear staining indicates that the presence of human nuclei in some of the fused myotube like, MHC positive structures (A,B). Quantitatively, human nuclear fusion index (C) and human MHC mRNA expression of B6 and C3 (D) are significantly greater than DSC+C2C12 group. In (D), y axis represents fold change related to DSC+C2C12 group. Scale bar, 50 µm.

Mentions: Upon co-culture with C2C12 cells, B6 and C3 also fused into multinucleated cells that expressed MHC within 1 wk (Fig. 4A,B). Human nuclear staining confirmed that multi-nucleated myotube structures contained both human and mouse nuclei (Fig. 4A,B). Upon co-culture of heterogeneous DSCs, 3 human cells (with positive human nuclear staining, HuNu) were identified among 1,475 cells, yielding a fusion index of 0.2% (Fig. 4C). In comparison, 52 human cells (with positive human nuclear staining, HuNu) were identified out of 1,551 cells when B6 clones were co-cultured with C2C12 cells, yielding a fusion index of 3.35% (Fig. 4C). Thus, the fusion index of myogenic clones was approximately 16–17 folds greater than their parent, heterogeneous stem cells. Furthermore, 51 human cells (with positive human nuclear staining, HuNu) were identified out of 1,482 cells when C3 clones were co-cultured with C2C12 cells, yielding a fusion index of 3.44% (Fig. 4C). Quantitative PCR revealed that co-culture of either B6 or C3 with C2C12 yielded significantly more human myosin heavy chain (MHC) mRNA than co-culture of heterogeneous DSCs with C2C12 (Fig. 4D).


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 upon co-culture with mouse skeletal myoblasts.Upon co-culture with mouse myoblast cell line (C2C12 cells) for 1 wk, some of the B6 and C3 cells again fused into multinucleated cells (A,B). Human nuclear staining indicates that the presence of human nuclei in some of the fused myotube like, MHC positive structures (A,B). Quantitatively, human nuclear fusion index (C) and human MHC mRNA expression of B6 and C3 (D) are significantly greater than DSC+C2C12 group. In (D), y axis represents fold change related to DSC+C2C12 group. Scale bar, 50 µm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2958121&req=5

pone-0013547-g004: Myogenic potential of clonal progeny of ectopic dental stem cells upon co-culture with mouse skeletal myoblasts.Upon co-culture with mouse myoblast cell line (C2C12 cells) for 1 wk, some of the B6 and C3 cells again fused into multinucleated cells (A,B). Human nuclear staining indicates that the presence of human nuclei in some of the fused myotube like, MHC positive structures (A,B). Quantitatively, human nuclear fusion index (C) and human MHC mRNA expression of B6 and C3 (D) are significantly greater than DSC+C2C12 group. In (D), y axis represents fold change related to DSC+C2C12 group. Scale bar, 50 µm.
Mentions: Upon co-culture with C2C12 cells, B6 and C3 also fused into multinucleated cells that expressed MHC within 1 wk (Fig. 4A,B). Human nuclear staining confirmed that multi-nucleated myotube structures contained both human and mouse nuclei (Fig. 4A,B). Upon co-culture of heterogeneous DSCs, 3 human cells (with positive human nuclear staining, HuNu) were identified among 1,475 cells, yielding a fusion index of 0.2% (Fig. 4C). In comparison, 52 human cells (with positive human nuclear staining, HuNu) were identified out of 1,551 cells when B6 clones were co-cultured with C2C12 cells, yielding a fusion index of 3.35% (Fig. 4C). Thus, the fusion index of myogenic clones was approximately 16–17 folds greater than their parent, heterogeneous stem cells. Furthermore, 51 human cells (with positive human nuclear staining, HuNu) were identified out of 1,482 cells when C3 clones were co-cultured with C2C12 cells, yielding a fusion index of 3.44% (Fig. 4C). Quantitative PCR revealed that co-culture of either B6 or C3 with C2C12 yielded significantly more human myosin heavy chain (MHC) mRNA than co-culture of heterogeneous DSCs with C2C12 (Fig. 4D).

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