Limits...
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.

Show MeSH

Related in: MedlinePlus

Characterization of heterogeneous stem/progenitor cells of the dental pulp.Immunostaining at passage 2 for Stro1(green) (A), CD146 (green) (B) and CD133 (red) (C) over DAPI-stained nuclei. Scale bars: 100 µm. D: Quantitative expression of CD146, Stro1 and CD133 at passages 2, 5 and 17 (N = 3).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2958121&req=5

pone-0013547-g001: Characterization of heterogeneous stem/progenitor cells of the dental pulp.Immunostaining at passage 2 for Stro1(green) (A), CD146 (green) (B) and CD133 (red) (C) over DAPI-stained nuclei. Scale bars: 100 µm. D: Quantitative expression of CD146, Stro1 and CD133 at passages 2, 5 and 17 (N = 3).

Mentions: Passage 2 (P2), heterogeneous DSCs expressed Stro1, CD146 and CD133 (Fig. 1A–C), but did not express CD34, CD44, CD105 or flk1 (data not shown). However, the number of Stro1, CD146 and CD133 positive cells decreased with passaging. At P5, few CD133+ cells remained, whereas Stro1+ and CD146+ cells decreased substantially (Fig. 1D). By P17, Stro1, CD133 and CD146 were virtually absent in heterogeneous cells (Fig. 1D). Heterogeneous DSCs continued to undergo population doubling (PD) during the tested 10 passages (Fig. 2A). The two isolated myogenic clones, B6 and C3, showed similar morphology and PD kinetics to their parent cell population (Fig. 2A) (without statistically significant differences in PD), indicating that the isolated clones by limiting dilution from heterogeneous cell populations were as capable of self renewal as their parent mononucleated and adherent cell populations.


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)

Characterization of heterogeneous stem/progenitor cells of the dental pulp.Immunostaining at passage 2 for Stro1(green) (A), CD146 (green) (B) and CD133 (red) (C) over DAPI-stained nuclei. Scale bars: 100 µm. D: Quantitative expression of CD146, Stro1 and CD133 at passages 2, 5 and 17 (N = 3).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2958121&req=5

pone-0013547-g001: Characterization of heterogeneous stem/progenitor cells of the dental pulp.Immunostaining at passage 2 for Stro1(green) (A), CD146 (green) (B) and CD133 (red) (C) over DAPI-stained nuclei. Scale bars: 100 µm. D: Quantitative expression of CD146, Stro1 and CD133 at passages 2, 5 and 17 (N = 3).
Mentions: Passage 2 (P2), heterogeneous DSCs expressed Stro1, CD146 and CD133 (Fig. 1A–C), but did not express CD34, CD44, CD105 or flk1 (data not shown). However, the number of Stro1, CD146 and CD133 positive cells decreased with passaging. At P5, few CD133+ cells remained, whereas Stro1+ and CD146+ cells decreased substantially (Fig. 1D). By P17, Stro1, CD133 and CD146 were virtually absent in heterogeneous cells (Fig. 1D). Heterogeneous DSCs continued to undergo population doubling (PD) during the tested 10 passages (Fig. 2A). The two isolated myogenic clones, B6 and C3, showed similar morphology and PD kinetics to their parent cell population (Fig. 2A) (without statistically significant differences in PD), indicating that the isolated clones by limiting dilution from heterogeneous cell populations were as capable of self renewal as their parent mononucleated and adherent cell populations.

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