A quantitative procedure for the dissociation of adult mammalian muscle into mononucleated cells

Yasin, Rose; Beers, Gisela Van; D, Bulien; Thompson, Edward J.

doi:10.1016/0014-4827(76)90056-2

Cited by 27 publications

(10 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…After removal of the fat and connective tissue, the remainder of the biopsy was minced into 1-mm3 pieces and digested with 0.1 % collagenase (Sigma, St. Louis, MO) and 0.1% trypsin (Gibco, Grand Island, NY) following a modification of the technique described by Yasin et a1. 44,45 Because the injection of fibroblasts into a muscle is harmful,' pure myoblasts were obtained by a dilution-cloning procedure. The clones were proliferated in MCDB 120 medium.'"…”

Section: Donor Muscle Biopsy and Myoblastmentioning

confidence: 99%

Human myoblast transplantation: Preliminary results of 4 cases

et al. 1992

View full text Add to dashboard Cite

Myoblasts from immunocompatible donors have been transplanted into the muscles (tibialis anterior, biceps brachii, and/or extensor carpi radialis longus) of 4 Duchenne patients in the advanced stages of the disease. Although no immunosuppressive treatment was used, none of the patients showed any clinical signs of rejection such as fever, redness, and inflammation. One patient transiently produced antibodies against the donor myoblasts as determined by cytofluorometric analysis. This patient and 2 others were shown to form antibodies against their donor's myotubes. Muscle biopsies of the injected tibialis anterior of 4 patients revealed that 80%, 75%, 25%, and 0% of the muscle fibers, respectively, showed some degree of dystrophin immunostaining. The contralateral noninjected muscles of the latter 3 patients did not contain any dystrophin positive fibers, while that of the first patient showed dystrophin expression in 16% of the fibers examined. Myoblasts were also injected into the extensor carpi radialis longus or the biceps brachii of these patients. A few months subsequent to injection, one patient was shown to have a 143% increase of strength during static wrist extension. This result must be interpreted with caution because a double-blind strength-measuring protocol was not used. Furthermore, we have noted that this change slowly decayed over time. The strength of 2 other patients was increased less remarkably (41% and 51%), while the strength of the fourth patient was unchanged.

show abstract

Section: Donor Muscle Biopsy and Myoblastmentioning

confidence: 99%

Human myoblast transplantation: Preliminary results of 4 cases

et al. 1992

View full text Add to dashboard Cite

show abstract

“…Alternatively, the cells that ultimately do form myotubes may not be capable of differentiating in vitro into mature fibers. If the defect is only expressed in the mature skeletal myotube, tissue culture would hardly reveal the abnormality A new cell technique, which involves tryptic digestion and mechanical dissociation (148,149), has been used successfully in the growth of muscle cells from a mononuclear cell suspension prepared from adult human skeletal muscle. This technique allows quantification of the number of mononuclear cells obtained per gram of tissue, as well as of plating eficiency (149).…”

Section: VIImentioning

confidence: 99%

Differentiation of skeletal muscle cells in culture.

Inestrosa

1982

Cell Struct. Funct.

View full text Add to dashboard Cite

“…Cultures of satellite cells from various species have been described; e.g., mouse (Kagawa et al, 1977), rat (Jones, 1977;Allen et al, 1984), human (Blau and Webster, 1981), chicken (Matsuda et al, 1983), and hamster (Yasin et al, 1976). However, direct quantitation of the number of satellite cells isolated was not possible in many of these studies because of the large amounts of myofibril fragments and cell debris in the final cell suspensions from both adult muscle (Young et al, 1978;Blau and Webster, 1981;Matsuda et al, 1983) and from muscles of older embryos (Hauschka, 1974).…”

Section: Introductionmentioning

confidence: 99%

Isolation and clonal analysis of satellite cells from chicken pectoralis muscle

Yablonka–Reuveni

Quinn

Nameroff

1987

Developmental Biology

150

View full text Add to dashboard Cite

Satellite cells, liberated from the breast muscle of young adult chickens by sequential treatment with collagenase and trypsin, were fractionated by Percoll density centrifugation to remove myofibril fragments and cell debris which otherwise heavily contaminate the preparation. This procedure allowed direct measurements of cell yields (1.5-4 X 10(5) cells/g tissue), plating efficiencies (27-40%) and identification of single cells in culture. In mass cultures, satellite cells gave rise to myotubes on the fifth day, and the progeny of these cells were sequentially passaged several times without losing myogenic traits. In clonal studies, over 90% of the satellite cells gave rise to large clones of which more than 99% were myogenic as demonstrated by the appearance of myotubes. The results obtained with satellite cells differ from observations made using embryonic muscle cell preparation from chicks. In the embryonic system massive formation of myotubes was observed following the third day of culture; sequential subculturing led to overgrowth of fibroblast-like cells following the first passage; and cells gave rise to both small myogenic clones (up to 16 terminally differentiated cells per clone) and non-myogenic clones in addition to large myogenic clones. We conclude that the isolated satellite cells represent a homogeneous cell population and reside in a stem cell compartment.

show abstract

A quantitative procedure for the dissociation of adult mammalian muscle into mononucleated cells

Cited by 27 publications

References 12 publications

Human myoblast transplantation: Preliminary results of 4 cases

Human myoblast transplantation: Preliminary results of 4 cases

Differentiation of skeletal muscle cells in culture.

Isolation and clonal analysis of satellite cells from chicken pectoralis muscle

Contact Info

Product

Resources

About