Stem cell antigen-1 regulates the tempo of muscle repair through effects on proliferation of α7 integrin-expressing myoblasts

Epting, Conrad L.; López, Javier E.; Pedersen, Anissa; Brown, Courtney; Spitz, Paul; Ursell, Philip C.; Bernstein, Harold S.

doi:10.1016/j.yexcr.2007.11.010

Cited by 28 publications

(33 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This new approach combined a 2-drops, purity mode with 8-coincidence logic with a negative selection step to abort contaminated large drops. A contaminated drop was identified by a nucleated cell signal expressing cell-surface markers for any one of multiple NMCs (CD31, CD45 and Sca-1) known to be present in muscle preparations 10,31,32 . A targeted drop was defined as a NMC-negative nucleated cell that labeled positively for cardiac Troponin T (cTnT), a sarcomeric regulatory protein only expressed in MCs.…”

Section: Resultsmentioning

confidence: 99%

Novel large-particle FACS purification of adult ventricular myocytes reveals accumulation of myosin and actin disproportionate to cell size and proteome in normal post-weaning development

López

Sharma

Ávila

et al. 2017

Journal of Molecular and Cellular Cardiology

Self Cite

View full text Add to dashboard Cite

Rationale Quantifying cellular proteins in ventricular myocytes (MCs) is challenging due to tissue heterogeneity and the variety of cell sizes in the heart. In post-weaning cardiac ontogeny, rod-shaped MCs make up the majority of the cardiac mass while remaining a minority of cardiac cells in number. Current biochemical analyses of cardiac proteins do not correlate well the content of MC-specific proteins to cell type or size in normally developing tissue. Objective To develop a new MC-specific large-particle fluorescent-activated cell sorting (LP-FACS) strategy for the purification of adult rod-shaped MCs. This approach is developed to enable growth-scaled measurements per-cell of the MC proteome and sarcomeric protein (i.e. myosin heavy chain (MyHC) and alpha-actin (α-actin)) content. Methods and Results Individual cardiac cells were isolated from 21-94 days old mice. An LP-FACS jet-in-air system with a 200-μm nozzle was defined by the first time to purify adult MCs. Cell-type specific immunophenotyping and sorting yielded ≥95% purity of adult MCs independently of cell morphology and size. This approach excluded other cell types and tissue contaminants from further analysis. MC proteome, MyHC and α-actin proteins were measured in linear biochemical assays normalized to cell numbers. Using the allometric coefficient α, we scaled the MC-specific rate of protein accumulation to growth post-weaning. MC-specific volumes (α=1.02) and global protein accumulation (α=0.94) were proportional (i.e. isometric) to body mass. In contrast, MyHC and α-actin accumulated at a much greater rate (i.e. hyperallometric) than body mass (α= 1.79 and 2.19 respectively) and MC volumes (α= 1.76 and 1.45 respectively). Conclusion Changes in MC proteome and cell volumes measured in LP-FACS purified MCs are proportional to body mass post-weaning. Oppositely, MyHC and α-actin are concentrated more rapidly than what would be expected from MC proteome accumulation, cell enlargement, or animal growth alone. LP-FACS provides a new standard for adult MC purification and an approach to scale the biochemical content of specific proteins or group of proteins per cell in enlarging MCs.

show abstract

Section: Resultsmentioning

confidence: 99%

Novel large-particle FACS purification of adult ventricular myocytes reveals accumulation of myosin and actin disproportionate to cell size and proteome in normal post-weaning development

López

Sharma

Ávila

et al. 2017

Journal of Molecular and Cellular Cardiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Moreover, treatment with TPPU in the MI animals resulted in a significant decrease in Thy pos cells compared with MI alone. Next, Ki67, a nuclear antigen that is expressed in actively cycling cells (26,27), was used to directly test the hypothesis that there is a significant increase in the proliferative capacity among CFs isolated from the MI model. There was a significant increase in the percentage of Ki67 in the Thy pos population in MI mice compared with sham animals.…”

Section: Effects Of Tppu On Cardiac Fibrosis In the Remote Zone In Thmentioning

confidence: 99%

Unique mechanistic insights into the beneficial effects of soluble epoxide hydrolase inhibitors in the prevention of cardiac fibrosis

Sirish

Liu

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

110

View full text Add to dashboard Cite

Tissue fibrosis represents one of the largest groups of diseases for which there are very few effective therapies. In the heart, myocardial infarction (MI) resulting in the loss of cardiac myocytes can culminate in adverse cardiac remodeling leading to eventual heart failure. Adverse cardiac remodeling includes myocyte hypertrophy, fibrosis, and electrical remodeling. We have previously demonstrated the beneficial effects of several potent soluble epoxide hydrolase inhibitors (sEHIs) in different models of cardiac hypertrophy and failure. Here, we directly determine the molecular mechanisms underlying the beneficial effects of sEHIs in cardiac remodeling post-MI. Treatment with a potent sEHI, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidine-4-yl)urea (TPPU), which was started 1 wk post-MI in a murine model, results in a significant improvement in cardiac function. Importantly, treatment with TPPU results in a decrease in cardiac fibrosis as quantified using histological and immunostaining techniques. Moreover, single-cell-based assays demonstrate that treatment with TPPU results in a significant decrease not only in the percentages but also the proliferative capacity of different populations of cardiac fibroblasts as well as a reduction in the migration of fibroblasts into the heart from the bone marrow. Our study provides evidence for a possible unique therapeutic strategy to reduce cardiac fibrosis and improve cardiac function post-MI.

show abstract

“…An unexpected finding was the loss of Sca-1 on L-MSCs 32 days after transplantation. Previous studies in skeletal myoblasts show that loss of Sca-1 is associated with reduced differentiation capacity [58]. Hence, future studies might address whether Sca-1 is a biomarker, or regulator of differentiation potential in L-MSCs.…”

mentioning

confidence: 95%

Lung-Derived Mesenchymal Stromal Cell Post-Transplantation Survival, Persistence, Paracrine Expression, and Repair of Elastase-Injured Lung

Hoffman

Paxson

Mazan

et al. 2011

Stem Cells and Development

View full text Add to dashboard Cite

While multipotent mesenchymal stromal cells have been recently isolated from adult lung (L-MSCs), there is very limited data on their biological properties and therapeutic potential in vivo. How L-MSCs compare with bone marrow-derived MSCs (BM-MSCs) is also unclear. In this study, we characterized L-MSC phenotype, clonogenicity, and differentiation potential, and compared L-MSCs to BM-MSCs in vivo survival, retention, paracrine gene expression, and repair or elastase injury after transplantation. L-MSCs were highly clonogenic, frequently expressed aldehyde dehydrogenase activity, and differentiated into osteocytes, chondrocytes, adipocytes, myofibroblasts, and smooth muscle cells. After intravenous injection (2 h), L-MSCs showed greater survival than BMMSCs; similarly, L-MSCs were significantly more resistant than BM-MSCs to anchorage independent culture (4 h) in vitro. Long after transplantation (4 or 32 days), a significantly higher number of CD45 neg L-MSCs were retained than BM-MSCs. By flow cytometry, L-MSCs expressed more intercellular adhesion molecule-1 (ICAM-1), platelet derived growth factor receptor alpha (PDGFRa), and integrin a2 than BM-MSCs; these proteins were found to modulate endothelial adherence, directional migration, and migration across Matrigel in L-MSCs. Further, L-MSCs with low ICAM-1 showed poorer lung retention and higher phagocytosis in vivo. Compared with BM-MSCs, L-MSCs expressed higher levels of several transcripts (e.g., Ccl2, Cxcl2, Cxcl10, IL-6, IL-11, Hgf, and Igf2) in vitro, although gene expression in vivo was increased by L-MSCs and BM-MSCs equivalently. Accordingly, both L-MSCs and BM-MSCs reduced elastase injury to the same extent. This study demonstrates that tissuespecific L-MSCs possess mechanisms that enhance their lung retention after intravenous transplantation, and produce substantial healing of elastase injury comparable to BM-MSCs.

show abstract

Stem cell antigen-1 regulates the tempo of muscle repair through effects on proliferation of α7 integrin-expressing myoblasts

Cited by 28 publications

References 53 publications

Novel large-particle FACS purification of adult ventricular myocytes reveals accumulation of myosin and actin disproportionate to cell size and proteome in normal post-weaning development

Novel large-particle FACS purification of adult ventricular myocytes reveals accumulation of myosin and actin disproportionate to cell size and proteome in normal post-weaning development

Unique mechanistic insights into the beneficial effects of soluble epoxide hydrolase inhibitors in the prevention of cardiac fibrosis

Lung-Derived Mesenchymal Stromal Cell Post-Transplantation Survival, Persistence, Paracrine Expression, and Repair of Elastase-Injured Lung

Contact Info

Product

Resources

About