Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Sep;71(9):3621–3625. doi: 10.1073/pnas.71.9.3621

The Use of Cytochalasin B to Distinguish Myoblasts from Fibroblasts in Cultures of Developing Chick Striated Muscle

J W Sanger 1,2
PMCID: PMC433827  PMID: 4530324

Abstract

Cytochalasin B (5 μg/ml) elicits a differential effect on myoblasts and fibroblasts in culture. After 1 day in culture in the presence of the drug, two types of cells were observed, round cells and cells with elongated arms, designated “arborized” cells. Both cell types were examined in the electron microscope. The round cells contained aggregates of thin and thick filaments as well as a few intact sarcomeres. Within the arms of the arborized cells were bundles of intermediate sized filaments (100 Å in diameter). The round cells could be shaken off the culture dish, washed free of cytochalasin B, and recultured to form myotubes. The remaining arborized cells lost their stellate shape when the drug was removed. The progeny of these cells gave rise to normal fibroblasts. Cytochalasin B, thus could be used to identify and isolate myoblasts prior to their fusion into developing muscle. It is suggested that this differential effect of the drug can be used to prepare pure cultures of fusible muscle cells uncontaminated by fibroblasts.

Keywords: electron microscopy, polarized light microscopy, birefringence, arborization

Full text

PDF
3621

Images in this article

3622Image
on p.3622
3623Image
on p.3623
3624Image
on p.3624
3624Image
on p.3624
3625Image
on p.3625

Selected References

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

  1. Abbott J., Schiltz J., Dienstman S., Holtzer H. The phenotypic complexity of myogenic clones. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1506–1510. doi: 10.1073/pnas.71.4.1506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carter S. B. Effects of cytochalasins on mammalian cells. Nature. 1967 Jan 21;213(5073):261–264. doi: 10.1038/213261a0. [DOI] [PubMed] [Google Scholar]
  3. Chlebowski J. S., Przbylski R. J., Cox P. G. Ultrastructural studies of lizard (Anolis carolinensis) myogenesis in vitro. Dev Biol. 1973 Jul;33(1):80–99. doi: 10.1016/0012-1606(73)90166-8. [DOI] [PubMed] [Google Scholar]
  4. Cox P. G. In vitro myogenesis of promuscle cells from the regenerating tail of the lizard, Anolis carolinensis. J Morphol. 1968 Sep;126(1):1–18. doi: 10.1002/jmor.1051260102. [DOI] [PubMed] [Google Scholar]
  5. Ebstensen R. D., Plagemann P. G. Cytochalasin B: inhibition of glucose and glucosamine transport. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1430–1434. doi: 10.1073/pnas.69.6.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Estensen R. D., Rosenberg M., Sheridan J. D., Wessells N. K., Spooner B. S., Ash J. F., Ludueña M. A., Wrenn J. T. Cytochalasin B: microfilaments and "contractile" processes. Science. 1971 Jul 23;173(3994):356–359. doi: 10.1126/science.173.3994.356. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Holtzer H., Sanger J. W. Cytochalasin-B: microfilaments, cell movement and what else? Dev Biol. 1972 Mar;27(3):444–446. [PubMed] [Google Scholar]
  9. Ishikawa H., Bischoff R., Holtzer H. Mitosis and intermediate-sized filaments in developing skeletal muscle. J Cell Biol. 1968 Sep;38(3):538–555. doi: 10.1083/jcb.38.3.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Manasek F. J., Burnside B., Stroman J. The sensitivity of developing cardiac myofibrils to cytochalasin-B (electron microscopy-polarized light-Z-bands-heartbeat). Proc Natl Acad Sci U S A. 1972 Feb;69(2):308–312. doi: 10.1073/pnas.69.2.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mayhew E., Maslow D. E. Cytochalasin B and the sialic acids of Ehrlich ascites cells. Exp Cell Res. 1974 Feb;83(2):255–260. doi: 10.1016/0014-4827(74)90337-1. [DOI] [PubMed] [Google Scholar]
  14. Mizel S. B. Differential effect of cytochalasin B on the two possible modes of 2-deoxyglucose transport in HeLa cells. Nat New Biol. 1973 May 23;243(125):125–126. [PubMed] [Google Scholar]
  15. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  16. Sachs H. G., McDonald T. F., Springer M. Cytochalasin B and embryonic heart muscle: contractility, excitability and ultrastructure. J Cell Sci. 1974 Jan;14(1):163–185. doi: 10.1242/jcs.14.1.163. [DOI] [PubMed] [Google Scholar]
  17. Sanger J. W., Holtzer H. Cytochalasin B: effects on cell morphology, cell adhesion, and mucopolysaccharide synthesis (cultured cells-contractile microfilaments-glycoproteins-embryonic cells-sorting-out). Proc Natl Acad Sci U S A. 1972 Jan;69(1):253–257. doi: 10.1073/pnas.69.1.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sanger J. W., Holtzer H. Cytochalasin-B: effects on cytokinesis, glycogen and 3 H-D-gluconse incorporation. Am J Anat. 1972 Oct;135(2):293–298. doi: 10.1002/aja.1001350214. [DOI] [PubMed] [Google Scholar]
  19. Sanger J. W., Holtzer S., Holtzer H. Effects of cytochalasin B on muscle cells in tissue culture. Nat New Biol. 1971 Jan 27;229(4):121–123. doi: 10.1038/newbio229121a0. [DOI] [PubMed] [Google Scholar]
  20. Wessells N. K., Spooner B. S., Ash J. F., Bradley M. O., Luduena M. A., Taylor E. L., Wrenn J. T., Yamada K. Microfilaments in cellular and developmental processes. Science. 1971 Jan 15;171(3967):135–143. doi: 10.1126/science.171.3967.135. [DOI] [PubMed] [Google Scholar]
  21. White N. K., Hauschka S. D. Muscle development in vitro. A new conditioned medium effect on colony differentiation. Exp Cell Res. 1971 Aug;67(2):479–482. doi: 10.1016/0014-4827(71)90437-x. [DOI] [PubMed] [Google Scholar]
  22. Zigmond S. H., Hirsch J. G. Cytochalasin B: inhibition of D-2-deoxyglucose transport into leukocytes and fibroblasts. Science. 1972 Jun 30;176(4042):1432–1434. doi: 10.1126/science.176.4042.1432. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES