Abstract
The maturation of striated muscle in primary cultures closely parallels the formation of striated muscle in vivo. Primary cultures thus serve as model systems for the study and manipulation of various aspects of muscle development including the regulation of gene expression, myofibril assembly, myocyte fusion, and myotube contraction. The following protocols provide instructions for the preparation of skeletal muscle cultures from either day 10 embryonic pectoralis muscle or day 2 somites and segmental plate mesoderm. Both methods will yield well-striated, multinucleated, contracting myotubes (Fig. 1) (1–4). Also included is a protocol for the preparation of cardiac cultures from day 7 embryonic heart. This method will yield well-striated, contracting cardiomyocytes (Fig. 1) (5,6).
An example of muscle prepared by each of the primary muscle culture protocols is illustrated in the following micrographs. Pectoralis muscle, segmental plate, and cardiac muscle cultures were immunofluorescently labeled with antibodies to α-actinin, MyoD, or titin, respectively. Multinucleated myotubes from embryonic pectoralis muscle cultures are shown in phase contrast (A) and with a sarcomeric distribution of α-actinin along striated myofibrils (B). Differentiating segmental plate cells contain MyoD in the nucleus (C). Striated cardiomyocytes from embryonic heart show titin with a sarcomeric localization (D).
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References
Lin, Z., Lu, M.-H., Schultheiss, T., Choi, J., Holtzer, H., DiLullo, C., et al. (1994) Sequential appearance of muscle-specific proteins in myoblasts as a function of time after cell division: evidence for a conserved myoblast differentiation program in skeletal muscle. Cell Motility Cytol. 29, 1–19.
Holtzer, H., Schultheiss, T., DiLullo, C., Choi, J., Costa, M., Lu, M.-H., and Holtzer, S. (1990) Autonomous expression of the differentiation programs of cells in the cardiac and skeletal myogenic lineages. Ann. NY Acad. Sci. 599, 158–169.
George-Weinstein, M., Gerhart, J., Foti, G., and Lash, J. W. (1994) Maturation of myogenic and chondrogenic cells in the presomitic mesoderm of the chick embryo. Exp. Cell Res. 211, 263–274.
George-Weinstein, M., Gerhart, J., Blitz, J., Simak, E., and Knudsen, K. (1997) N-cadherin promotes the commitment and differentiation of skeletal muscle precursor cells. Dev. Biol. 185, 14–24.
Lu, M.-H., DiLullo, C., Schultheiss, T., Holtzer, S., Murray, J. M., Choi, J., et al. (1992) The vinculin/sarcomeric-α-actinin/α-actin nexus in cultured cardiac myocytes. J. Cell Biol. 117, 1007–1022.
Dlugosz, A. A., Antin, P. B., Nachmias, V. T., and Holtzer, H. (1984) The relationship between stress fiber-like structures and nascent myofibrils in cultured cardiac myocytes. J. Cell Biol. 99, 2268–2278.
Hamburger, V. and Hamilton, H. L. (1951) A series of normal stages in development of the chick embryo. J. Morphol. 88, 49–92.
George-Weinstein, M., Gerhart, J., Reed, R., Flynn, J., Callihan, B., Mattiacci, M., et al. (1996) Skeletal myogenesis: The preferred pathway of chick embryo epiblast cells in vitro. Dev. Biol. 173, 279–291.
Ahrens, P. B., Solursh, M., and Reiter, R. S. (1977) Stage related capacity for limb chondrogenesis in cell culture. Dev. Biol. 60, 69–82.
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© 2000 Humana Press Inc., Totowa, NJ
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Di Lullo, C., Gerhart, J., George-Weinstein, M. (2000). Preparation of Chick Striated Muscle Cultures. In: Tuan, R.S., Lo, C.W. (eds) Developmental Biology Protocols. Methods in Molecular Biology™, vol 137. Humana Press. https://doi.org/10.1385/1-59259-066-7:337
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DOI: https://doi.org/10.1385/1-59259-066-7:337
Publisher Name: Humana Press
Print ISBN: 978-0-89603-854-7
Online ISBN: 978-1-59259-066-7
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