Creatine kinase isoenzymes in cultured human muscle cells. II. A study of carrier females for Duchenne muscular dystrophy by needle and open biopsy

Cavanagh, N. P. C.; Franklin, Gillian I.; Hughes, B.R.; Yasin, Rose; Phillips, Elsa; Beers, Gisela Van; Thompson, Edward J.

doi:10.1016/0009-8981(81)90075-9

Cited by 8 publications

(1 citation statement)

References 10 publications

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“…When muscle is dissociated and the cells plated in culture, both myoblasts and fibroblasts are obtained and the rate of proliferation of the fibroblasts frequently exceeds that of the myoblasts. DMD muscle cultures have been compared with normal muscle cultures and a number of differences reported, including increased collagen synthesis (2), decreased specific activity of creatine kinase (3), increased ratio of nonmuscle to muscle creatine kinase isozymes (3-5), altered lipid biosynthesis (6), and clustered myotube morphology (5,7,8). However, it is unclear whether these differences are intrinsic to the DMD myoblasts and related to the disease or a reflection of the proportion of fibroblasts present in the mixed cultures analyzed.…”

mentioning

confidence: 99%

Defective myoblasts identified in Duchenne muscular dystrophy.

Blau¹,

Webster²,

Pavlath³

1983

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

293

199

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A defect in the proliferative capacity of satellite cells, mononucleated precursors of mature muscle fibers, was found in clonal analyses of cells cultured from Duchenne muscular dystrophy (DMD) patients. The total yield of myoblasts per gram of muscle biopsy was decreased to 5% of normal. Of the DMD myoblast clones obtained, a large proportion -contained a morphological class of flat distended cells that had an increased generation time and ceased to proliferate beyond 100-1,000 cells but could be induced to fuse and form myotubes. The altered muscle phenotype was detected in all cultures from DMD patients but was rarely found among myoblasts of controls. By age 14 yr, it comprised as many as 90% of DMD myoblasts. The remaining DMD myoblast clones, which initially grew well, had severely impaired proliferative capacity upon passage and further cultivation. Eventually all myoblasts from DMD muscle tissue exhibited defective growth potential. In contrast, the fibroblast yield and proliferative capacity from DMD samples did not differ from normal. Based on these findings, we propose a hypothesis for the etiology of DMD: Dividing myoblasts are required for muscle growth and maintenance, and the limited capacity of DMD myoblasts to grow is directly related to the progressive muscle degeneration characteristic of the disease.Duchenne muscular dystrophy (DMD) is a chronic, lethal Xlinked disease that affects 1 in 4,800 males (1). Although DMD is well defined clinically, the cellular and biochemical basis for the disease is unknown. A major problem in the biological study of muscular dystrophy has been the heterogeneous nature of muscle tissue. Muscle is composed of a mixture of cell types present in variable ratios, especially in disease states. When muscle is dissociated and the cells plated in culture, both myoblasts and fibroblasts are obtained and the rate of proliferation of the fibroblasts frequently exceeds that of the myoblasts. DMD muscle cultures have been compared with normal muscle cultures and a number of differences reported, including increased collagen synthesis (2), decreased specific activity of creatine kinase (3), increased ratio of nonmuscle to muscle creatine kinase isozymes (3-5), altered lipid biosynthesis (6), and clustered myotube morphology (5,7,8). However, it is unclear whether these differences are intrinsic to the DMD myoblasts and related to the disease or a reflection of the proportion of fibroblasts present in the mixed cultures analyzed.We have developed methods for obtaining pure populations of human myoblasts that can be studied under standardized conditions (9). Our methods use postnatal muscle as a source.Consequently, the myoblasts obtained in culture are muscle satellite cells, the mononucleated myoblasts of mature muscle fibers, positioned between the basement membrane and the sarcolemma (10). It is these satellite cells that are capable of proliferating in vitro (11, 12) and are probably responsible for postnatal muscle regeneration and fiber growth in vivo. We have...

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mentioning

confidence: 99%

Defective myoblasts identified in Duchenne muscular dystrophy.

Blau¹,

Webster²,

Pavlath³

1983

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

293

199

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Tissue culture studies of muscle disorders: Part 1. Techniques, cell growth, morphology, cell surface

Witkowski¹

1986

Muscle and Nerve

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Tissue culture has been used extensively in studies of human inherited disorders, and its application in the field of the neuromuscular disorders has increased rapidly in recent years. This review, covering the period 1977 to 1984, deals with tissue culture studies of both human and animal muscle disorders, although Duchenne muscular dystrophy (DMD) figures prominently because of the overwhelming interest in that disorder. The review is in two parts. In the first part, I discuss technical innovations in the field, the morphology and growth of cells, and a variety of studies related to the cell surface. Important findings in relation to DMD include reports of abnormal growth rates and reduced lifespan of DMD cells, hypersensitivity to DNA-damaging agents, abnormal cell-to-cell and cell-to-substratum adhesion, and a more "fluid" cell membrane. However, these findings are controversial or have so far been reported only by single laboratories.

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Tissue culture studies of muscle disorders: Part 2. Biochemical studies, nerve‐muscle culture, metabolic myopathies, and animal models

Witkowski¹

1986

Muscle and Nerve

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This review continues with studies of protein, lipid, and purine metabolism of Duchenne muscular dystrophy (DMD) cells in vitro and of muscle cells in combined culture with nerve cells. In vitro studies of human metabolic myopathies are tabulated. Results using the hamster, chicken, and mouse (dy25, dy, mdg, and mdx) myopathies are discussed. Interesting findings include suggestions of altered collagen synthesis by DMD cells. Analysis of cell proteins by two-dimensional gel electrophoresis and the use of combined nerve-muscle cultures remain important areas of development. It is disappointing that so few attempts have been made to repeat significant findings in this field, and when a number of laboratories have examined the same phenomenon, the results are often contradictory. It remains to be shown how these various abnormalities found in cells in vitro are related to each other and to those pathologic features of diseased muscle observed in vivo.

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Creatine kinase isoenzymes in cultured human muscle cells. II. A study of carrier females for Duchenne muscular dystrophy by needle and open biopsy

Cited by 8 publications

References 10 publications

Defective myoblasts identified in Duchenne muscular dystrophy.

Defective myoblasts identified in Duchenne muscular dystrophy.

Tissue culture studies of muscle disorders: Part 1. Techniques, cell growth, morphology, cell surface

Tissue culture studies of muscle disorders: Part 2. Biochemical studies, nerve‐muscle culture, metabolic myopathies, and animal models

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