Assaying Human Myogenic Progenitor Cell Activity by Reconstitution of Muscle Fibers and Satellite Cells in Immunodeficient Mice

Introduction Treatment options for critical limb ischaemia (CLI) are limited. Recent evidence has suggested that even with successful revascularisation, patients often show little functional improvement. This has been attributed to a musculopathy that occurs in CLI. Myogenic progenitor satellite cells (SCs) provide skeletal muscle with an intrinsic ability to regenerate. It has been shown that there is an increase in SCs in ischaemic muscle, however their function in ischaemia is poorly understood and we hypothesize that ischaemia has a detrimental effect on SC function. Methods Gastrocnemius muscle biopsies were taken from CLI patients and compared with non ischaemic control biopsies. The phenotypical changes and frequency of satellite cells were investigated using PAX 7 immunohistochemistry and western blot. C2C12 myoblasts were used in vitro, to investigate the effect of ischaemia on muscle progenitor cell function. Myoblasts were exposed to simulated ischaemia for 24, 48 and 72hrs. Proliferation rates were assessed using an MTT assay. Differentiation and apoptosis were assessed by MYOD and cleaved caspase 3 western blotting respectively. Results There is an increased expression of PAX 7 in CLI muscle biopsies, shown by both immunostaining and western blot analysis, suggesting an increased number of SCs in ischaemic human skeletal muscle (p<0.05). Myoblasts cultured in ischaemic conditions demonstrated decreased cell proliferation, reduced myogenic differentiation (decreased MYOD expression), and increased apoptosis (increased cleaved caspase 3 expression). Conclusion Despite an upregulation of SCs in ischaemic tissue, their function is suppressed in ischaemic conditions and this may be contributing to the poor functional recovery of patients post revascularisation. Enhancement of muscle regeneration in ischaemia may be a useful therapeutic adjunct in the treatment of CLI.

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Hematopoietic Stem Cells Can Give Rise to Satellite-Like Cells in Skeletal Muscles.

Blood ◽

10.1182/blood.v104.11.2690.2690 ◽

2004 ◽

Vol 104 (11) ◽

pp. 2690-2690

Author(s):

Momoko Yoshimoto ◽

Toshio Heike ◽

Mitsutaka Shiota ◽

Hirohiko Kobayashi ◽

Katsutsugu Umeda ◽

...

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Basal Lamina ◽

Satellite Cells ◽

Time Course ◽

Mononuclear Cells ◽

Muscle Fibers ◽

Cell Activity ◽

Fish Analysis ◽

Hematopoietic Stem

Abstract Recent studies have indicated that bone marrow cells can regenerate damaged muscles, but also that they can adopt phenotype of other cells by cell fusion. It has also been reported that single hematopoietic stem cells (HSCs) can regenerate skeletal muscle although it is still controversial whether HSCs differentiate into satellite cells in muscle or not. In order to investigate the roles of HSCs in muscle regeneration and whether they can generate satellite cells or not, we purified and injected CD45+Lin−Sca-1+c-kit+(CD45+KSL) HSCs labeled by green fluorescent protein (GFP) into mice with or without irradiation. We examined time-course behavior of HSCs in recipient muscles with a fluorescent stereomicroscope and then immunohitochemical staining during the early and late phase after transplantation. Our direct visualization system gave evidence of massive GFP signals in all the muscles of only irradiated mice in early phase after transplantation. Transverse cryostat sections showed GFP+ Myosin+ muscle fibers along with numerous GFP+ hematopoietic cells in damaged muscle. We also found myogenin+GFP+ cells like myoblasts in very low number. These phenomena were temporal and GFP signals had dramatically reduced 30 days after transplantation. FISH analysis confirmed the GFP-DNAs in the nuclei of muscle fibers. These results suggested that most of GFP+HSCs fused with myofibers and participated in regeneration of damaged muscles, and a very few HSCs can differentiate into myoblast like cells expressing myogenin. After 6 months, GFP+ fibers could be hardly detected but GFP+c-Met+ mononuclear cells were located beneath the laminin+ basal lamina. Single fiber cultures from these mice showed proliferation of GFP+ fibers. These results suggested that HSC-derived cells settled beneath the basal lamina like satellite cells and might acquired the satellite cell activity.

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Satellite cell activation in the stretch-enlarged anterior latissimus dorsi muscle of the adult quail

AJP Cell Physiology ◽

10.1152/ajpcell.1991.260.2.c206 ◽

1991 ◽

Vol 260 (2) ◽

pp. C206-C212 ◽

Cited By ~ 48

Author(s):

P. K. Winchester ◽

M. E. Davis ◽

S. E. Alway ◽

W. J. Gonyea

Keyword(s):

Satellite Cell ◽

Satellite Cells ◽

Latissimus Dorsi ◽

Tritiated Thymidine ◽

Cell Activity ◽

Muscle Length ◽

Latissimus Dorsi Muscle ◽

Cell Labeling ◽

Dorsi Muscle ◽

Anterior Latissimus Dorsi

Satellite cell activity was examined in the stretch-enlarge anterior latissimus dorsi muscle (ALD) of the adult quail. Thirty-seven birds had a weight equal to 10% of their body mass attached to one wing while the contralateral wing served as an intra-animal control. At various time intervals after application of the wing weight (from 1 to 30 days), the birds were injected with tritiated thymidine and killed 1 h later. Stretched muscle length was greater by day 1 and mass by day 3 when compared with the contralateral muscle. Satellite cells actively synthesizing DNA were quantitated in fiber segments of the control and stretched ALD. A minimum of 1,500 muscle nuclei (satellite cell nuclei and myonuclei) were counted in each muscle. Labeling in stretched muscle was expressed by the percent labeled nuclei per total nuclei counted. Satellite cell labeling was initiated by day 1, peaked between days 3 and 7, and was not statistically different from control values at day 30. These results demonstrate that satellite cells are induced to enter the cell cycle in the stretch-enlarged ALD muscle from the adult quail, and the peak of proliferative activity is within the first week of stretch.

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Muscle Fibers in Regenerating Crayfish Motor Nerves

Journal of Neurophysiology ◽

10.1152/jn.1997.78.6.3498 ◽

1997 ◽

Vol 78 (6) ◽

pp. 3498-3501 ◽

Cited By ~ 4

Author(s):

Joanne Pearce ◽

Kristin M. Krause ◽

C. K. Govind

Keyword(s):

Signaling Pathway ◽

Satellite Cells ◽

Blood Cells ◽

Axon Regeneration ◽

Muscle Fibers ◽

Nerve Terminals ◽

Motor Axons ◽

The Third ◽

Motor Nerves ◽

Regenerating Nerve

Pearce, Joanne, Kristin M. Krause, and C. K. Govind. Muscle fibers in regenerating crayfish motor nerves. J. Neurophysiol. 78: 3498–3501, 1997. Single discrete muscle fibers were found in regenerating motor nerves in adult crayfish. The regenerating nerves were from native or transplanted ganglia in the third abdominal segments and consisted of several motor axons. The proximal end of these motor axons showed numerous sprouts. Muscle fibers in these regenerating nerves appeared newly developed and were innervated by excitatory nerve terminals. A likely source of these novel muscle fibers may be blood cells in the nerve or satellite cells from neighboring muscle. Contacts made by axon sprouts with other axon sprouts, glia, and muscle fiber, in the form of a dense bar with clustered clear vesicles, characterized the regenerating nerve. These contacts may provide a possible signaling pathway for axon regeneration and myogenesis.

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Leucine/glutamic acid/lysine protein 1 is localized to subsets of myonuclei in bovine muscle fibers and satellite cells

Journal of Animal Science ◽

10.2527/jas.2009-1998 ◽

2009 ◽

Vol 87 (10) ◽

pp. 3134-3141 ◽

Cited By ~ 3

Author(s):

S. E. Ouellette ◽

J. Li ◽

W. Sun ◽

S. Tsuda ◽

D. K. Walker ◽

...

Keyword(s):

Glutamic Acid ◽

Satellite Cells ◽

Muscle Fibers ◽

Bovine Muscle

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Understanding the Role of Antiviral Cytokines and Chemokines on Neural Stem/Progenitor Cell Activity and Survival

Viral Immunology ◽

10.1089/vim.2018.0091 ◽

2019 ◽

Vol 32 (1) ◽

pp. 15-24 ◽

Cited By ~ 3

Author(s):

Manisha N. Chandwani ◽

Patrick S. Creisher ◽

Lauren A. O'Donnell

Keyword(s):

Progenitor Cell ◽

Cell Activity ◽

Cytokines And Chemokines

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Skeletal muscle regeneration is altered in the R6/2 mouse model of Huntington's disease

10.1101/2022.01.11.475914 ◽

2022 ◽

Author(s):

Sanzana Hoque ◽

Marie Sjogren ◽

Valerie Allamand ◽

Kinga Gawlik ◽

Naomi Franke ◽

...

Keyword(s):

Skeletal Muscle ◽

Huntington's Disease ◽

Huntington’S Disease ◽

Muscle Damage ◽

Satellite Cell ◽

Mouse Models ◽

Satellite Cells ◽

Muscle Fibers ◽

Cag Repeat ◽

Skeletal Muscle Regeneration

Huntington's disease (HD) is caused by CAG repeat expansion in the huntingtin (HTT) gene. Skeletal muscle wasting alongside central pathology is a well-recognized phenomenon seen in patients with HD and HD mouse models. HD muscle atrophy progresses with disease and affects prognosis and quality of life. Satellite cells, progenitors of mature skeletal muscle fibers, are essential for proliferation, differentiation, and repair of muscle tissue in response to muscle injury or exercise. In this study, we aim to investigate the effect of mutant HTT on the differentiation and regeneration capacity of HD muscle by employing in vitro mononuclear skeletal muscle cell isolation and in vivo acute muscle damage model in R6/2 mice. We found that, similar to R6/2 adult mice, neonatal R6/2 mice also exhibit a significant reduction in myofiber width and morphological changes in gastrocnemius and soleus muscles compared to WT mice. Cardiotoxin (CTX)-induced acute muscle damage in R6/2 and WT mice showed that the Pax7+ satellite cell pool was dampened in R6/2 mice at 4 weeks post-injection, and R6/2 mice exhibited an altered inflammatory profile in response to acute damage. Our results suggest that, in addition to the mutant HTT degenerative effects in mature muscle fibers, expression of mutant HTT in satellite cells might alter developmental and regenerative processes to contribute to the progressive muscle mass loss in HD. Taken together, the results presented here encourage further studies evaluating the underlying mechanisms of satellite cell dysfunction in HD mouse models.

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Nfatc1’s Role in Mammary Epithelial Morphogenesis and Basal Stem/progenitor Cell Self-renewal

Journal of Mammary Gland Biology and Neoplasia ◽

10.1007/s10911-021-09502-6 ◽

2021 ◽

Author(s):

Melissa McNeil ◽

Yingying Han ◽

Peng Sun ◽

Kazuhide Watanabe ◽

Jun Jiang ◽

...

Keyword(s):

Stem Cell ◽

Mammary Gland ◽

Progenitor Cell ◽

Cell Function ◽

Cell Activity ◽

Basal Layer ◽

Mammary Epithelial ◽

Epithelial Morphogenesis ◽

Small Subset ◽

Self Renewal

AbstractMammary gland is an outstanding system to study the regulatory mechanisms governing adult epithelial stem cell activity. Stem cells in the basal layer of the mammary gland fuel the morphogenesis and regeneration of a complex epithelial network during development and upon transplantation. The self-renewal of basal stem/progenitor cells is subjected to regulation by both cell-intrinsic and extrinsic mechanisms. Nfatc1 is a transcription factor that regulates breast tumorigenesis and metastasis, but its role in mammary epithelial development and stem cell function has not been investigated. Here we show that Nfatc1 is expressed in a small subset of mammary basal epithelial cells and its epithelial-specific deletion results in mild defects in side branching and basal-luminal cell balance. Moreover, Nfatc1-deficient basal cells exhibit reduced colony forming ability in vitro and somewhat compromised regenerative potential upon transplantation. Thus, our study provides evidence for a detectable yet non-essential role of Nfatc1 in mammary epithelial morphogenesis and basal stem/progenitor cell self-renewal.

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