Abstract
An assessment of the replicative life-span of myoblasts is of fundamental importance in designing treatment strategies for Duchenne muscular dystrophy (DMD) based on cell or gene therapy. To ascertain myoblast life-span, or the total number of cell divisions of which a myoblast was capable, we serially passaged and counted the progeny of individual myoblasts until they senesced. We compared the life-span of myoblasts from eight DMD patients with controls: three individuals with no known neuromuscular disease, three DMD carriers, and three patients with other muscle degenerative diseases. A decline in replicative capacity was observed with increasing donor age, which was markedly accelerated for DMD relative to control myoblasts. The average myoblast from a 5-year-old control was capable of 56 doublings, or a potential yield of approximately 1017 cells per cell. By contrast, at 2 years of age, the typical age at clinical onset, only 6% of DMD myoblasts had a life-span of 50 doublings in tissue culture, and by age 7 DMD myoblasts capable of 10 doublings were rare. Our results suggest that the myoblasts (satellite cells) of even the youngest DMD patients have undergone extensive division in an attempt to regenerate degenerating myofibers. These findings have implications for therapeutic intervention in DMD involving genetic engineering and myoblast implantation.
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Literature cited
Kunkel, L.A., Monaco, A.P., Middlesworth, W., Ochs, H.D., and Latt, S.A. (1985).Proc. Natl. Acad. Sci. USA 824778–4782.
Liechti-Gallati, S., Koenig, M., Kunkel, L.M., Frey, D., Boltshauser, E., Schneider, V., Braga, S. and, Moser, H. (1989).Hum. Genet. 81343–348.
Ray, P.N., Belfall, B., Duff, C., Logan, C., Kean, V., Thompson, M.W., Sylvester, J.E., Gorski, J.L., Schmickel, R.D., and Worton, R.G. (1985).Nature 318672–675.
Webster, C., Silberstein, L., Hays, A.P., and Blau, H.M. (1988).Cell 32503–513.
Bradley, W.G., Budgson, P., Larson, P.F., Papapetropoulos, T.A., and Jenkison, M. (1972).J. Neurol. Neurosurg. Psych. 35451–455.
Pearson, C.M. (1962).Brain 85109–126.
Morizumi, H., Hizawa, K., Nunomura, S., and Ii, K. (1984).Acta Pathol. Jpn. 341221–1242.
Watkins, S.C., and Cullen, M.J. (1985).Neuropathol. Appl. Neurobiol. 11447–460.
Schmalbruch, H. (1984).Neurology 3460–65.
Lipton, B.H. (1979). InMuscle Regeneration (ed.) Mauro, A. (Raven Press, New York), pp. 31–40.
Ontell, M., Feng, K.C., Klueber, K., Dunn, R.F., and Taylor, F. (1984).Anat. Rec. 208159–174.
Zaks, S.I., and Sheff, M.F. (1982).Muscle Nerve 5152–161.
Cullen, M.J., and Watkins, S.C. (1981).Adv. Physiol. Sci. 24341–349.
Wakayama, Y., and Schotland, D.L. (1979). InMuscle Regeneration (ed.) Mauro, A. (Raven Press, New York), pp. 121–129.
Hellmuth, A.E., and Allbrook, D. (1973). InBasic Research in Myology, Part 1, (ed.) Kakulas, B.A. (Excerpta Medica, Amsterdam), pp. 343–345.
Moss, F.P., and Leblond, C.P. (1971).Anat. Rec. 170421–436.
Aloisis, M., Mussini, I., and Schiaggino, S. (1973). InBasic Research in Myology, Part 1, (ed.) Kakulas, B.A. (Excerpta Medica, Amsterdam), pp. 338–342.
Carlson, B.M. (1973).Am. J. Anat. 137119–150.
Webster, C., Filippi, G., Rinaldi, A., Mastropaolo, C., Tondi, M., Siniscalco, M., and Blau, H.M. (1986).Hum. Genet. 7474–80.
Blau, H.M., and Webster, C. (1981).Proc. Natl. Acad. Sci. U.S.A. 785623–5627.
Chiu, C.-P., and Blau, H.M. (1984).Cell 37879–887.
Walsh, F.S., Dickson, G., Moore, S.E., and Barton, C.H. (1989).Nature 339516.
Webster, C., Pavlath, G.K., Parks, D.R., Walsh, F.S., and Blau, H.M. (1987).Exp. Cell Res. 174252–265.
Blau, H.M., Webster, C., and Pavlath, G.K. (1983).Proc. Natl. Acad. Sci. U.S.A. 804856–4860.
Hauschka, S.D., Linkhart, T.A., Clegg, C., and Merrill, G. (1979). InMuscle Regeneration (ed.) Mauro, A. (Raven Press, New York), pp. 311–322.
Hayflick, L. (1965).Exp. Cell Res. 37614–636.
Rheinwald, J.G., and Green, H. (1975).Cell 6331–344.
Hudgson, P., Pearce, G.W., and Walton, J.N. (1967).Brain 90565–582.
Dubowitz, V., and Brooke, M.H. (1973).Muscle Biopsy: A Modern Approach. Major Problems in Neurology Monograph Series, Vol. 2 (W.B. Saunders, Philadelphia).
Blau, H.M., Webster, C., Chiu, C.-P., Guttman, S., and Chandler, F. (1983).Exp. Cell Res. 144495–502.
Ionasescu, V., and Ionasescu, R. (1982).J. Neurol. Sci. 5479–87.
Yasin, R., Kundy, D., and Thompson, E.J. (1982).Exp. Cell Res. 138419–422.
Hauschka, S.D. (1974).Dev. Biol. 37345–368.
Cossu, G., Cicinelli, P., Fieri, C., Coletta, M., and Molinaro, M. (1985).Exp. Cell Res. 180402–411.
Miller, J.B., and Stockdale, F.E. (1986).J. Cell Biol. 1032197–2208.
Toutant, M., Montarras, D., and Fiszman, M.Y. (1984).Exp. Biol. Med. 910–15.
White, N.K., Bonner, P.H., Nelson, D.R., and Hauschka, S.D. (1975).Dev. Biol. 44346–361.
Ontell, M. (1979). InMuscle Regeneration (ed.) Mauro, A. (Raven Press, New York), pp. 137–146.
Wright, W.E. (1985).Exp. Cell Res. 157343–354.
Koenig, M., Hoffman, E.P., Bertelson, C.J., Monaco, A.P., Feener, C., and Kunkel, L.M. (1987).Cell 50509–517.
Feener, C.A., Koenig, M., and Kunkel, L.M. (1989).Nature 338509–511.
Partridge, T.A., Morgan, J.E., Coulton, G.R., Hoffman, E.P., and Kunkel, L.M. (1989).Nature 337176–179.
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Webster, C., Blau, H.M. Accelerated age-related decline in replicative life-span of Duchenne muscular dystrophy myoblasts: Implications for cell and gene therapy. Somat Cell Mol Genet 16, 557–565 (1990). https://doi.org/10.1007/BF01233096
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DOI: https://doi.org/10.1007/BF01233096