scholarly journals Evidence for a myogenic stem cell that is exhausted in dystrophic muscle

2000 ◽  
Vol 113 (12) ◽  
pp. 2299-2308 ◽  
Author(s):  
L. Heslop ◽  
J.E. Morgan ◽  
T.A. Partridge
Keyword(s):  
Skeletal Muscles ◽  
Precursor Cells ◽  
Mdx Mouse ◽  
Muscle Fibres ◽  
Significant Extent ◽  
Dystrophic Muscle ◽  
Myogenic Cells ◽  
Mouse Muscle ◽  
Muscle Precursor ◽  
Muscle Precursor Cells

Injection of the myotoxin notexin, was found to induce regeneration in muscles that had been subjected to 18 Gy of radiation. This finding was unexpected as irradiation doses of this magnitude are known to block regeneration in dystrophic (mdx) mouse muscle. To investigate this phenomenon further we subjected mdx and normal (C57Bl/10) muscle to irradiation and notexin treatment and analysed them in two ways. First by counting the number of newly regenerated myofibres expressing developmental myosin in cryosections of damaged muscles. Second, by isolating single myofibres from treated muscles and counting the number of muscle precursor cells issuing from these over 2 day and 5 day periods. After irradiation neither normal nor dystrophic muscles regenerate to any significant extent. Moreover, single myofibres cultured from such muscles produce very few muscle precursor cells and these undergo little or no proliferation. However, when irradiated normal and mdx muscles were subsequently treated with notexin, regeneration was observed. In addition, some of the single myofibres produced rapidly proliferative muscle precursor cells when cultured. This occurred more frequently, and the myogenic cells proliferated more extensively, with fibres cultured from normal compared with dystrophic muscles. Even after 25 Gy, notexin induced some regeneration but no proliferative myogenic cells remained associated with the muscle fibres. Thus, skeletal muscles contain a number of functionally distinct populations of myogenic cells. Most are radiation sensitive. However, some survive 18 Gy as proliferative myogenic cells that can be evoked by extreme conditions of muscle damage; this population is markedly diminished in muscles of the mdx mouse. A small third population survives 25 Gy and forms muscle but not proliferative myogenic cells.

An evaluation of cell separation techniques in a model mixed cell population

1992 ◽  
Vol 102 (4) ◽  
pp. 789-798
Author(s):  
S.J. Murphy ◽  
D.J. Watt ◽  
G.E. Jones
Keyword(s):  
Cell Population ◽  
Cell Separation ◽  
Separation Efficiency ◽  
Neuronal Cell ◽  
Quantitative Measure ◽  
Precursor Cells ◽  
Muscle Fibres ◽  
Myogenic Cells ◽  
Muscle Precursor ◽  
Muscle Precursor Cells

Muscle precursor cells may act not only as a means of inserting normal genes into diseased muscle fibres, in order to correct or alleviate a genetically inherited myopathy, but recent demonstrations have shown they may prove an invaluable tool for the expression of, and systemic dissemination of, non-muscle gene products. If muscle precursor cells are proved to act as such widespread vectors in terms of gene therapy, then it is imperative that methods are properly elucidated to produce large populations of pure viable myogenic cells for such purposes. In the past, many methods of cell separation have been investigated but carry with them the problems of either a lack of myogenic purity of the population or poor percentage recovery of the original cell population. In the present work we have investigated two methods for segregating myogenic from non-myogenic cells and have critically reviewed the efficiency of separation of the two techniques used. To obtain a quantitative measure of separation efficiency, segregation was carried out on a 1:1 mixture of murine C2 myogenic and murine 3T3 fibroblastic cells. To distinguish between C2 and 3T3 cells, the latter were prelabelled with the fluorescent strain carboxyfluorescein diacetate succinimyl ester (CFSE). Once incorporated into the cell, CFSE remains there, thus preventing transfer of the label to C2 cells. Both methods of separation used depend on the affinity of myogenic cells for the monoclonal antibody Mab H28, which specifically binds to the mouse neuronal cell adhesion molecule N-CAM, but differ in that one method, “panning”, completes segregation by adherence of N-CAM positive cells to a dish precoated with secondary IgG antibody whereas in the other separation proceeds by the use of commercially available IgG-coated magnetic beads. Results indicate magnetic bead separation to be more efficient than panning if the beads are precoated with 0.1% gelatin.

Human Muscle Precursor Cells Form Human-Derived Myofibers in Skeletal Muscles of Nonhuman Primates: A Potential New Preclinical Setting to Test Myogenic Cells of Human Origin for Cell Therapy of Myopathies

2020 ◽  
Vol 79 (12) ◽  
pp. 1265-1275
Author(s):  
Daniel Skuk ◽  
Jacques P Tremblay
Keyword(s):  
Muscle Regeneration ◽  
De Novo ◽  
Lymphocytic Infiltration ◽  
Precursor Cells ◽  
Human Muscle ◽  
Myogenic Cells ◽  
Cell Therapies ◽  
Muscle Precursor ◽  
Muscle Precursor Cells

Abstract This study aimed to verify if human myogenic cells could participate in muscle regeneration in macaques. This experimental setting would grant researchers a model that could better evaluate the effects of cell therapies in myopathies with a better translation to human patients. Human muscle precursor cells (MPCs) were cultured in vitro and transduced with ß-galactosidase. The cells were subsequently injected into 1-cm3 muscle regions of 6 macaques immunosuppressed with tacrolimus and dexamethasone. Allogeneic ß-galactosidase+ MPCs were injected in other regions as positive controls. Some cell-grafted regions were electroporated to induce extensive muscle regeneration. MPC-grafted regions were sampled 1 month later and analyzed by histology. There were ß-galactosidase+ myofibers in both the regions grafted with human and macaque MPCs. Electroporation increased the engraftment of human MPCs in the same way as in macaque allografts. The histological analysis (hematoxylin and eosin, CD8, and CD4 immunodetection) demonstrated an absence of cellular rejection in most MPC-grafted regions, as well as minimal lymphocytic infiltration in the regions transplanted with human MPCs in the individual with the lowest tacrolimus levels. Circulating de novo anti-donor antibodies were not detected. In conclusion, we report the successful engraftment of human myogenic cells in macaques, which was possible using tacrolimus-based immunosuppression.

scholarly journals Heterogeneity among muscle precursor cells in adult skeletal muscles with differing regenerative capacities

1998 ◽  
Vol 212 (4) ◽  
pp. 495-508 ◽  
Author(s):  
Grace K. Pavlath ◽  
Deepa Thaloor ◽  
Thomas A. Rando ◽  
Monica Cheong ◽  
Arthur W. English ◽  
...  
Keyword(s):  
Skeletal Muscles ◽  
Precursor Cells ◽  
Muscle Precursor ◽  
Muscle Precursor Cells

scholarly journals The Drosophila FUS ortholog cabeza promotes adult founder myoblast selection by Xrp1-dependent regulation of FGF signaling

2020 ◽  
Author(s):  
Marica Catinozzi ◽  
Moushami Mallik ◽  
Marie Frickenhaus ◽  
Marije Been ◽  
Céline Sijlmans ◽  
...  
Keyword(s):  
Dna Binding ◽  
Skeletal Muscles ◽  
Precursor Cells ◽  
Abdominal Muscles ◽  
Cell Selection ◽  
Fgf Signaling ◽  
Fgf Receptor ◽  
Muscle Precursor ◽  
Founder Cells ◽  
Muscle Precursor Cells

AbstractThe number of adult myofibers in Drosophila is determined by the number of founder myoblasts selected from a myoblast pool, a process governed by fibroblast growth factor (FGF) signaling. Here, we show that loss of cabeza (caz) function results in a reduced number of adult founder myoblasts, leading to a reduced number and misorientation of adult dorsal abdominal muscles. Genetic experiments revealed that loss of caz function in both adult myoblasts and neurons contributes to caz mutant muscle phenotypes. Selective overexpression of the FGF receptor Htl or the FGF receptor-specific signaling molecule Stumps in adult myoblasts partially rescued caz mutant muscle phenotypes, and Stumps levels were reduced in caz mutant founder myoblasts, indicating FGF pathway deregulation. In both adult myoblasts and neurons, caz mutant muscle phenotypes were mediated by increased expression levels of Xrp1, a DNA-binding protein involved in gene expression regulation. Xrp1-induced phenotypes were dependent on the DNA-binding capacity of its AT-hook motif, and increased Xrp1 levels in founder myoblasts reduced Stumps expression. Thus, control of Xrp1 expression by Caz is required for regulation of Stumps expression in founder myoblasts, resulting in correct founder myoblast selection.Author SummarySkeletal muscles mediate movement, and therefore, proper structure and function of skeletal muscles is required for respiration, locomotion, and posture. Adult muscles arise from fusion of muscle precursor cells during development. In the fruit fly Drosophila melanogaster, muscle precursor cells come in two flavors: founder cells and fusion-competent cells. The number of founder cells selected during development corresponds to the number of adult muscles formed. Here, we report that inactivation of the Drosophila caz gene results in muscle developmental defects. Loss of caz function in both muscle precursor cells and the nerve cells that innervate muscles contributes to the muscle developmental defect. At the molecular level, loss of caz function leads to increased levels of Xrp1. Xrp1 regulates the expression of many other genes, including genes that produce components of the FGF signaling pathway, which is known to be involved in founder cell selection. In all, we uncovered a novel molecular mechanism that regulates founder cell selection during muscle development.

scholarly journals Dermal fibroblasts convert to a myogenic lineage in mdx mouse muscle

1995 ◽  
Vol 108 (1) ◽  
pp. 207-214 ◽  
Author(s):  
A.J. Gibson ◽  
J. Karasinski ◽  
J. Relvas ◽  
J. Moss ◽  
T.G. Sherratt ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Muscle Protein ◽  
Dermal Fibroblasts ◽  
Muscle Disease ◽  
Mdx Mouse ◽  
Muscle Fibres ◽  
Normal Muscle ◽  
Myogenic Cells ◽  
Mouse Muscle

Duchenne muscular dystrophy is a primary muscle disease that manifests itself in young boys as a result of a defect in a gene located on the X-chromosome. This gene codes for dystrophin, a normal muscle protein that is located beneath the sarcolemma of muscle fibres. Therapies to alleviate this disease have centred on implanting normal muscle precursor cells into dystrophic fibres to compensate for the lack of this gene and its product. To date, donor cells for implantation in such therapy have been of myogenic origin, derived from paternal biopsies. Success in human muscle, however, has been limited and may reflect immune rejection problems. To overcome this problem the patient's own myogenic cells, with the dystrophin gene inserted, could be used, but this could lead to other problems, since these cells are those that are functionally compromised by the disease. Here, we report the presence of high numbers of dystrophin-positive fibres after implanting dermal fibroblasts from normal mice into the muscle of the mdx mouse-the genetic homologue of Duchenne muscular dystrophy. Dystrophin-positive fibres were also abundant in mdx muscle following the implantation of cloned dermal fibroblasts from the normal mouse. Our results suggest the in vivo conversion of these non-myogenic cells to the myogenic pathway resulting in the formation of dystrophin-positive muscle fibres in the deficient host. The use of dermal fibroblasts may provide an alternative approach to the previously attempted myoblast transfer therapy, which in human trials has yielded disappointing results.

Implanted myoblasts not only fuse with myofibers but also survive as muscle precursor cells

1993 ◽  
Vol 105 (4) ◽  
pp. 957-963 ◽  
Author(s):  
S.N. Yao ◽  
K. Kurachi
Keyword(s):  
Gene Transfer ◽  
Muscle Regeneration ◽  
Precursor Cells ◽  
Additional Mechanism ◽  
Myogenic Cells ◽  
Muscle Precursor ◽  
Primary Myoblasts ◽  
Muscle Precursor Cells

Intramuscular implanted myoblasts can fuse with existing myofibers. Here we report that implanted primary myoblasts marked with retroviral transgenes can also persist as muscle precursor cells. These cells can be recovered as viable myoblasts from muscles of recipient mice even months after myoblast implantation, and they can fully resume expression of the transgenes in culture. Upon re-implantation into muscles, they again not only fuse with existing myofibers, but also survive as muscle precursor cells in the tissue. These reserve myogenic cells should be able to contribute to host myofibers in muscle regeneration when the recombinant myofibers are damaged, providing an additional mechanism to maintain a persistent expression of transgenes delivered by myoblast-mediated gene transfer.

506: Restoration of Innervated Urinary Sphincter Function Using Muscle Precursor Cells

2004 ◽  
Vol 171 (4S) ◽  
pp. 135-135
Author(s):  
Rene Yiou ◽  
James J. Yoo ◽  
Anthony Atala
Keyword(s):  
Precursor Cells ◽  
Sphincter Function ◽  
Muscle Precursor ◽  
Urinary Sphincter ◽  
Muscle Precursor Cells
Sign in / Sign up

Export Citation Format

Share Document