Production of Microcytospheres

  • Gerd G. Maul
  • Josef Weibel


Cytoplasmic vesicles can be isolated by several methods which utilize agents with cross-linking capabilities, formaldehyde or glutaraldehyde (Scott et al., 1979). These partially fixed vesicles are unable to attach to a substrate. Since, in certain types of experiments, cytoplasmic vesicles containing only the active ribosomal components and a normal cell membrane are desirable, a method was needed to ensure minimal contamination of vesicles with other organelles. Cytochalasin B has been used in the past to break down the microfilaments associated with the cell membrane (Carter, 1967) and to enucleate cells (Prescott et al., 1971; Veomett et al., 1976; Wigler and Weinstein, 1975; Gopalakrishnan and Tompson, 1975). The resulting cyto-plasts can then be fused with cells or karyoplasts of different origin (Shay, 1977). However, these cytoplasts contain all of the cellular organelles. We describe here a procedure that is speedy, sterile, and applicable to a variety of experiments in which it is essential that cytoplasmic parts of one cell type containing only defined components are fused with other cells. These types of cytoplasmic vesicles are named microcytospheres.


HeLa Cell Mitotic Cell Size Marker Plastic Dish Cytoplasmic Vesicle 
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  1. Carter, S. B., 1967, Effects of cytochalasins on mammalian cells, Nature 213:261–264.PubMedCrossRefGoogle Scholar
  2. Clark, M. A., and Shay, J. W., 1982, Long lived cytoplasmicfactors that suppress adrenal steroidogenesis, Proc. Natl. Acad. Sci USA 79:1144–1148.PubMedCrossRefGoogle Scholar
  3. Gopalakrishnan, T. V., and Tompson, E. B., 1975, A method for enucleating cultured mammalian cells, Exp. Cell Res. 96:435–439.PubMedCrossRefGoogle Scholar
  4. Maul, G. G., Maul, H. M., Scogna, J. E., Lieberman, M. W., Stein, G. S., Hsu, B. Y., and Borun, T. W., 1972, Time sequence of nuclear pore formation in phytohemagglutinin-stimulated lymphocytes and in HeLa cells during the cell cycle, J. Cell. Biol. 55:433–447.PubMedCrossRefGoogle Scholar
  5. Prescott, D. M., Myerson, D., and Wallace, J., 1971, Enucleation of mammalian cells with cytochalasin B, Exp. Cell Res. 71:480–485.CrossRefGoogle Scholar
  6. Scott, R. E., and Maercklein, P. B., 1979, Plasma membrane vesiculation in 3T3 and SV3T3 cells: Factors affecting the process of vesiculation, J. Cell Sci. 35:245–252.PubMedGoogle Scholar
  7. Scott, R. E., Perkins, R. G., Zschunke, M. A., Hoerl, B. J., and Maercklein, P. B., 1979, Plasma membrane vesiculation in 3T3 and SV3T3 cells: Morphological and biochemical characterization, J. Cell Sci. 35:229–243.PubMedGoogle Scholar
  8. Shay, J. W., 1977, Selection of reconstituted cells from karyoplasts fused to chloramphenicol-resistant cytoplasts, Proc. Natl. Acad. Sci. USA 74:2461–2464.PubMedCrossRefGoogle Scholar
  9. Sunkara, P. S., Al-Bader, A. A., and Rao, P. N., 1977, Mitoplasts: Mitotic cells minus the chromosomes, Exp. Cell Res. 107:444–447.PubMedCrossRefGoogle Scholar
  10. Veomett, G., Shay, J. W., Hough, P. V., and Prescott, D. M., 1976, Large-scale enucleation of mammalian cells, in: Methods in Cell Biology, Volume XIII (D. M. Prescott, ed.), Academic Press, New York, pp. 1–6.Google Scholar
  11. Wigler, M. H., and Weinstein, J. B., 1975, A preparative method for obtaining enucleated mammalian cells, Biochem. Biophys. Res. Commun. 63:669–674.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • Gerd G. Maul
    • 1
  • Josef Weibel
    • 1
  1. 1.Wistar Institute of Anatomy and BiologyPhiladelphiaUSA

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