Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1972 Jan 1;52(1):52–65. doi: 10.1083/jcb.52.1.52

A KINETIC ANALYSIS OF MYOGENESIS IN VITRO

Michael C O'Neill 1, Frank E Stockdale 1
PMCID: PMC2108680  PMID: 5006948

Abstract

Conditions which yielded reproducible growth kinetics with extensive, relatively synchronous differentiation are described for chick muscle cultures. The effects of cell density and medium changes on the timing of cell fusion were examined. Low-density cultures which received a change of medium at 24 hr after plating show the highest rate of cell fusion, increasing from 15 to 80% fused cells in a 10 hr period. These optimal culture conditions were employed to reexamine two questions from the earlier literature on muscle culture: (a) can cells which normally would fuse at the end of one cell cycle be forced to go through another cell cycle before fusion; and (b) how soon after its final S period can a cell complete fusion? In answer to the first question, it was found that if the medium is changed, many cells which would otherwise fuse can be made to undergo another cell cycle before fusion. In the second case, radioautographs were made from cultures incubated with tritiated thymidine for various times at the beginning of the fusion period. These show labeled nuclei in myotubes as early as 3 hr after the beginning of the incubation period. This indicates that cells can fuse as early as the beginning of the G1 period, and suggests that there is not an obligatory exit from the cell cycle or a prolonged G1 period before cell fusion and differentiation during myogenesis.

Full Text

The Full Text of this article is available as a PDF (827.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bischoff R., Holtzer H. Mitosis and the processes of differentiation of myogenic cells in vitro. J Cell Biol. 1969 Apr;41(1):188–200. doi: 10.1083/jcb.41.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. HERRMANN H., WHITE B. N., COOPER M. The accumulation of tissue components in the leg muscle of the developing chick. J Cell Physiol. 1957 Apr;49(2):227–252. doi: 10.1002/jcp.1030490206. [DOI] [PubMed] [Google Scholar]
  3. Hauschka S. D., Konigsberg I. R. The influence of collagen on the development of muscle clones. Proc Natl Acad Sci U S A. 1966 Jan;55(1):119–126. doi: 10.1073/pnas.55.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. KECK K. An ultramicro technique for the determination of deoxypentose nucleic acid. Arch Biochem Biophys. 1956 Aug;63(2):446–451. doi: 10.1016/0003-9861(56)90059-5. [DOI] [PubMed] [Google Scholar]
  5. KONIGSBERG I. R. Clonal analysis of myogenesis. Science. 1963 Jun 21;140(3573):1273–1284. doi: 10.1126/science.140.3573.1273. [DOI] [PubMed] [Google Scholar]
  6. Kaighn M. E., Ebert J. D., Stott P. M. The susceptibility of differentiating muscle clones to Rous sarcoma virus. Proc Natl Acad Sci U S A. 1966 Jul;56(1):133–140. doi: 10.1073/pnas.56.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kruse P. F., Jr, Miedema E. Production and characterization of multiple-layered populations of animal cells. J Cell Biol. 1965 Nov;27(2):273–279. doi: 10.1083/jcb.27.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LASH J. W., HOLTZER H., SWIFT H. Regeneration of mature skeletal muscle. Anat Rec. 1957 Aug;128(4):679–697. doi: 10.1002/ar.1091280404. [DOI] [PubMed] [Google Scholar]
  9. Marchok A. C., Herrmann H. Studies of muscle development. I. Changes in cell proliferation. Dev Biol. 1967 Feb;15(2):129–155. doi: 10.1016/0012-1606(67)90010-3. [DOI] [PubMed] [Google Scholar]
  10. O'Neill M., Strohman R. C. Changes in DNA polymerase activity associated with cell fusion in cultures of embryonic muscle. J Cell Physiol. 1969 Feb;73(1):61–68. doi: 10.1002/jcp.1040730109. [DOI] [PubMed] [Google Scholar]
  11. Okazaki K., Holtzer H. Myogenesis: fusion, myosin synthesis, and the mitotic cycle. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1484–1490. doi: 10.1073/pnas.56.5.1484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. STOCKDALE F. E., HOLTZER H. DNA synthesis and myogenesis. Exp Cell Res. 1961 Sep;24:508–520. doi: 10.1016/0014-4827(61)90450-5. [DOI] [PubMed] [Google Scholar]
  13. Smets L. A. Discrepancies between precursor uptake and DNA synthesis in mammalian cells. J Cell Physiol. 1969 Aug;74(1):63–66. doi: 10.1002/jcp.1040740109. [DOI] [PubMed] [Google Scholar]
  14. Todaro G. J., Lazar G. K., Green H. The initiation of cell division in a contact-inhibited mammalian cell line. J Cell Physiol. 1965 Dec;66(3):325–333. doi: 10.1002/jcp.1030660310. [DOI] [PubMed] [Google Scholar]
  15. Yaffe D. Developmental changes preceding cell fusion during muscle differentiation in vitro. Exp Cell Res. 1971 May;66(1):33–48. doi: 10.1016/s0014-4827(71)80008-3. [DOI] [PubMed] [Google Scholar]
  16. Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci U S A. 1968 Oct;61(2):477–483. doi: 10.1073/pnas.61.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. de Turenne M., Daillie J. Utilisation de la thymidine exogene par la glande sericigene du ver a soie incubee in vitro. I. Compartimentage des nucleotides intracellulaires d'origine exogene. Biochim Biophys Acta. 1969 Aug 20;186(2):267–279. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES