Regeneration of adult newt skeletal muscle tissue in vitro

Development ◽  
1983 ◽  
Vol 77 (1) ◽  
pp. 255-271
Author(s):  
Joan A. Schrag ◽  
Jo Ann Cameron
Keyword(s):  
Skeletal Muscle ◽  
Primary Cultures ◽  
Muscle Fibres ◽  
Myogenic Cells ◽  
Subsequent Formation ◽  
Myotube Formation ◽  
Adult Newt ◽  
Muscle Cultures ◽  
Minced Muscle

Explants and cells of forelimb muscle from adult Notophthalmus viridescens were cultured for periods up to 160 days in MEM-based medium supplemented with serum, hormones, andantibiotics. Explants which were not minced prior to culture contained muscle fibres withhealthy myonuclei and no evidence of dedifferentiation after four weeks. Explants which were minced prior to culture contained degenerated muscle fibres after 1 day and no evidence of dedifferentiation after four weeks. Mononucleated cells from both minced and non-minced explants proliferated. Cell proliferation and myotube formation was greater in the minced muscle cultures. Proliferation and fusion of myoblasts and subsequent formation ofmyofibrils were observed on the plate in primary cultures. Secondarily transferred cells proliferated and fused into myotubes. Although adult newt muscle does not contain satellite cells, myogenesis in this amphibian followed the same course as all other vertebrate skeletal muscle: proliferation of mononucleated myogenic cells, fusion of the myoblaststo form syncytia, and eventual accumulation of myofibrils. The ultimate source of the myogenic cells was not identified; however, the absence of dedifferentiation of the mature fibres and the occurrence of myogenesis in cultures of minced muscle explants demonstratedthat the regenerated muscle developed from a population of mononucleated cells whose origin did not depend upon dedifferentiation of intact fibres.

scholarly journals Purified lectin from skeletal muscle inhibits myotube formation in vitro.

1980 ◽  
Vol 85 (3) ◽  
pp. 617-625 ◽  
Author(s):  
R G MacBride ◽  
R J Przybylski
Keyword(s):  
Skeletal Muscle ◽  
Chick Embryo ◽  
Culture Medium ◽  
Horse Serum ◽  
Pectoral Muscle ◽  
Embryonic Chick ◽  
Myotube Formation ◽  
Chick Embryo Extract ◽  
Muscle Cultures

A lactose-extractable lectin obtained from 14--16-d embryonic chick pectoral muscle and myotube muscle cultures by affinity chromatography inhibited myotube formation in culture. When applied to muscle cultures at 0.09 micrograms/ml, the purified lectin produced variable effects on the inhibition of myotube formation related to the time and length of application, suggesting that components of the culture medium and/or temperature produced inactivation. Hemagglutination assays showed that the lectin was inactivated by horse serum and by chick embryo extract but not by L-15 salt solution at 4 degrees C. Incubation in L-15 solution at 37 degrees C with or without 2 mM dithiothreitol resulted in inactivation in 2--3 h. To maximize the effect of the lectin on the inhibition of myotube formation, primary muscle cultures were grown in low [Ca+2] medium to inhibit fusion, and then [Ca+2] was increased to elicit fusion in the absence and presence of lectin with solution renewal every 2 h. Without lectin, myotube formation was normal, whereas, with lectin, it was inhibited by 93%. Continued incubation at 37 degrees C. without renewal of lectin resulted in myotube formation, suggesting reversibility by lectin inactivation.

Reversible suppression of skeletal myotube formation in vitro obtained by varying [CO2]

1979 ◽  
Vol 237 (3) ◽  
pp. C166-C176 ◽  
Author(s):  
R. J. Przybylski ◽  
J. C. Bullaro ◽  
R. MacBride
Keyword(s):  
Skeletal Muscle ◽  
Cell Proliferation ◽  
Primary Cultures ◽  
Myoblast Differentiation ◽  
Cell Recognition ◽  
Myotube Formation ◽  
Low Co2 ◽  
Cytoplasmic Structures ◽  
Chicken Skeletal Muscle

In primary cultures of chicken skeletal muscle, decreasing the [CO2] of the gaseous phase below 10(-3)% resulted in inhibition of cell proliferation and cytolysis. With 10(-3)% CO2-air, cell proliferation was slightly retarded and myotube formation was inhibited approximately 90% compared to cultures receiving 5% CO2-air. Changes in pH were not effective. Culture in low [CO2] resulted in the accumulation of lipoidal inclusions and unique cytoplasmic structures. Increasing time in culture with low [CO2] resulted in an increase in the length of G1 of the cell cycle. The inhibition was reversed by the addition of 5% CO2-air at any time in culture up to 2 wk with a minimum time of 3--6 h required. Lipoidal inclusions decreased in number and the unique cytoplasmic structures were absent. During the first 3 days in culture, myoblasts showing dependence on [CO2] for myotube formation increased in number, and the effect of elevated [CO2] on these cells was long lasting. The data suggest that some aspect of myoblast differentiation relating to cell recognition and fusion is affected by decreased [CO2].

scholarly journals Recycled algae-based carbon materials as electroconductive 3D printed skeletal muscle tissue engineering scaffolds

2021 ◽  
Vol 32 (7) ◽  
Author(s):  
Selva Bilge ◽  
Emre Ergene ◽  
Ebru Talak ◽  
Seyda Gokyer ◽  
Yusuf Osman Donar ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Tissue Engineering ◽  
Electrical Stimulation ◽  
Muscle Tissue ◽  
Carbonaceous Material ◽  
Hydrothermal Carbonization ◽  
Skeletal Muscle Tissue ◽  
Myotube Formation ◽  
3D Printed

AbstractSkeletal muscle is an electrically and mechanically active tissue that contains highly oriented, densely packed myofibrils. The tissue has self-regeneration capacity upon injury, which is limited in the cases of volumetric muscle loss. Several regenerative therapies have been developed in order to enhance this capacity, as well as to structurally and mechanically support the defect site during regeneration. Among them, biomimetic approaches that recapitulate the native microenvironment of the tissue in terms of parallel-aligned structure and biophysical signals were shown to be effective. In this study, we have developed 3D printed aligned and electrically active scaffolds in which the electrical conductivity was provided by carbonaceous material (CM) derived from algae-based biomass. The synthesis of this conductive and functional CM consisted of eco-friendly synthesis procedure such as pre-carbonization and multi-walled carbon nanotube (MWCNT) catalysis. CM obtained from biomass via hydrothermal carbonization (CM-03) and its ash form (CM-03K) were doped within poly(ɛ-caprolactone) (PCL) matrix and 3D printed to form scaffolds with aligned fibers for structural biomimicry. Scaffolds were seeded with C2C12 mouse myoblasts and subjected to electrical stimulation during the in vitro culture. Enhanced myotube formation was observed in electroactive groups compared to their non-conductive counterparts and it was observed that myotube formation and myotube maturity were significantly increased for CM-03 group after electrical stimulation. The results have therefore showed that the CM obtained from macroalgae biomass is a promising novel source for the production of the electrically conductive scaffolds for skeletal muscle tissue engineering.

scholarly journals Interleukin 4 Moderately Affects Competence of Pluripotent Stem Cells for Myogenic Conversion

2019 ◽  
Vol 20 (16) ◽  
pp. 3932 ◽  
Author(s):  
Barbara Świerczek-Lasek ◽  
Jacek Neska ◽  
Agata Kominek ◽  
Łukasz Tolak ◽  
Tomasz Czajkowski ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Pluripotent Stem Cells ◽  
Muscle Development ◽  
Embryonic Stem ◽  
Interleukin 4 ◽  
Myogenic Cells ◽  
The Impact

Pluripotent stem cells convert into skeletal muscle tissue during teratoma formation or chimeric animal development. Thus, they are characterized by naive myogenic potential. Numerous attempts have been made to develop protocols enabling efficient and safe conversion of pluripotent stem cells into functional myogenic cells in vitro. Despite significant progress in the field, generation of myogenic cells from pluripotent stem cells is still challenging—i.e., currently available methods require genetic modifications, animal-derived reagents, or are long lasting—and, therefore, should be further improved. In the current study, we investigated the influence of interleukin 4, a factor regulating inter alia migration and fusion of myogenic cells and necessary for proper skeletal muscle development and maintenance, on pluripotent stem cells. We assessed the impact of interleukin 4 on proliferation, selected gene expression, and ability to fuse in case of both undifferentiated and differentiating mouse embryonic stem cells. Our results revealed that interleukin 4 slightly improves fusion of pluripotent stem cells with myoblasts leading to the formation of hybrid myotubes. Moreover, it increases the level of early myogenic genes such as Mesogenin1, Pax3, and Pax7 in differentiating embryonic stem cells. Thus, interleukin 4 moderately enhances competence of mouse pluripotent stem cells for myogenic conversion.

Altered sodium current response to intracellular fatty acids in halothane-hypersensitive skeletal muscle

1996 ◽  
Vol 271 (1) ◽  
pp. C347-C353 ◽  
Author(s):  
S. J. Wieland ◽  
Q. H. Gong ◽  
J. E. Fletcher ◽  
H. Rosenberg
Keyword(s):  
Skeletal Muscle ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Sodium Current ◽  
Primary Cultures ◽  
Na Channel ◽  
Current Response ◽  
Whole Cell ◽  
Na Currents

Biopsies of human skeletal muscle were analyzed by an in vitro contracture test (IVCT) for responsiveness to a halothane challenge: noncontracting (nonresponsive; IVCT-) and contracting (IVCT+). A muscle biopsy that is IVCT+ indicates potential malignant hyperthermia (MH) susceptibility. Primary cultures were grown from portions of the skeletal muscle biopsies, and voltage-activated currents were measured by whole cell recording in the presence or absence of 2-5 microM intracellular arachidonic or oleic acids. In untreated IVCT- cells, Na+ currents were predominantly tetrodotoxin (TTX) insensitive, indicating that most of the current was carried through the embryonic SkM2 isoform of the Na+ channel. Inclusion of fatty acids in the recording pipette of IVCT- cells produced an increase in voltage-activated Na+ currents during 20 min of recording. Approximately 70% of currents in fatty acid-treated cells were TTX sensitive, indicating activation of the adult SkM1 isoform of the Na+ channel. In contrast to IVCT- cells, IVCT+ cells expressed Na+ currents that were predominantly TTX sensitive even in the absence of added fatty acid, thus showing a relatively large baseline functional expression of SkM1 channels. Addition of fatty acids to the recording pipette produced little further change in the magnitude or TTX sensitivity of the whole cell currents in IVCT+ cells, suggesting altered functional regulation of Na+ channels in MH muscle.

Spontaneous myogenic differentiation of Flk-1-positive cells from adult pancreas and other nonmuscle tissues

2008 ◽  
Vol 294 (2) ◽  
pp. C604-C612 ◽  
Author(s):  
Giuliana Di Rocco ◽  
Alessandra Tritarelli ◽  
Gabriele Toietta ◽  
Ilaria Gatto ◽  
Maria Grazia Iachininoto ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Myogenic Differentiation ◽  
Small Subset ◽  
Myogenic Cells ◽  
Adult Mice ◽  
Primary Myoblasts ◽  
Thymic Medulla ◽  
First Time

At the embryonic or fetal stages, autonomously myogenic cells (AMCs), i.e., cells able to spontaneously differentiate into skeletal myotubes, have been identified from several different sites other than skeletal muscle, including the vascular compartment. However, in the adult animal, AMCs from skeletal muscle-devoid tissues have been described in only two cases. One is represented by thymic myoid cells, a restricted population of committed myogenic progenitors of unknown derivation present in the thymic medulla; the other is represented by a small subset of adipose tissue-associated cells, which we recently identified. In the present study we report, for the first time, the presence of spontaneously differentiating myogenic precursors in the pancreas and in other skeletal muscle-devoid organs such as spleen and stomach, as well as in the periaortic tissue of adult mice. Immunomagnetic selection procedures indicate that AMCs derive from Flk-1+ progenitors. Individual clones of myogenic cells from nonmuscle organs are morphologically and functionally indistinguishable from skeletal muscle-derived primary myoblasts. Moreover, they can be induced to proliferate in vitro and are able to participate in muscle regeneration in vivo. Thus, we provide evidence that fully competent myogenic progenitors can be derived from the Flk-1+ compartment of several adult tissues that are embryologically unrelated to skeletal muscle.

scholarly journals Skeletal Myogenic Progenitors Originating from Embryonic Dorsal Aorta Coexpress Endothelial and Myogenic Markers and Contribute to Postnatal Muscle Growth and Regeneration

1999 ◽  
Vol 147 (4) ◽  
pp. 869-878 ◽  
Author(s):  
Luciana De Angelis ◽  
Libera Berghella ◽  
Marcello Coletta ◽  
Laura Lattanzi ◽  
Malvina Zanchi ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Satellite Cells ◽  
Developmental Stages ◽  
Vascular System ◽  
Muscle Growth ◽  
Paraxial Mesoderm ◽  
Dorsal Aorta ◽  
Myogenic Cells ◽  
Adult Skeletal Muscle

Skeletal muscle in vertebrates is derived from somites, epithelial structures of the paraxial mesoderm, yet many unrelated reports describe the occasional appearance of myogenic cells from tissues of nonsomite origin, suggesting either transdifferentiation or the persistence of a multipotent progenitor. Here, we show that clonable skeletal myogenic cells are present in the embryonic dorsal aorta of mouse embryos. This finding is based on a detailed clonal analysis of different tissue anlagen at various developmental stages. In vitro, these myogenic cells show the same morphology as satellite cells derived from adult skeletal muscle, and express a number of myogenic and endothelial markers. Surprisingly, the latter are also expressed by adult satellite cells. Furthermore, it is possible to clone myogenic cells from limbs of mutant c-Met−/− embryos, which lack appendicular muscles, but have a normal vascular system. Upon transplantation, aorta-derived myogenic cells participate in postnatal muscle growth and regeneration, and fuse with resident satellite cells. The potential of the vascular system to generate skeletal muscle cells may explain observations of nonsomite skeletal myogenesis and raises the possibility that a subset of satellite cells may derive from the vascular system.

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