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Nuclear Transplantation with Mammalian Cells

  • Margaret J. Hightower
  • Joseph J. Lucas

Abstract

Early microsurgical nuclear transplantation experiments with nonmammalian systems suggested that cytoplasmic elements participate in the regulation of nuclear gene expression and replication. With cells from Rana pipiens (Briggs and King, 1960), Xenopus leavis (Gurdon, 1962), and Drosophila melanogaster (Okada et al., 1974), it was shown that egg cell cytoplasm could redirect the differentiative pathway of nuclei from cells at much later stages of development. Moreover, in Stentor coeruleus, for example, nuclear DNA synthesis likewise appeared to be regulated, at least in part, by cytoplasmic factors (deTerra, 1967). With mammalian cells, numerous somatic cell hybridization experiments demonstrated that the patterns of gene expression of two parental cell types could be stably altered when a hybrid cell containing a mixed genome was constructed [reviewed by Ringertz and Savage (1976); Lucas (1982)]. For example, rat hepatoma cells that secreted albumin were fused to mouse fibroblasts that did not. Some hybrid clones secreted only mouse or rat albumin, while others secreted both rat and mouse albumin (Peterson and Weiss, 1972). Results of this and other similar experiments showed that a complex array of interactions is possible when two very different cell types are fused. They also suggested the existence, in animal cells, of elements which can interact functionally with a foreign nucleus and either positively or negatively regulate the expression of certain genes.

Keywords

Nuclear Transplantation Sendai Virus Normal Growth Medium Cytoplasmic Fragment Cell BioI 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Arndt-Jovin, D. J., and Jovin, T. M., 1977, Analysis and sorting of living cells according to deoxyribonucleic acid content, J. Histochem. Cytochem. 25:585–589.PubMedCrossRefGoogle Scholar
  2. Boyum, A., 1968, Isolation of mononuclear cells and granulocytes from human blood, Scand. J. Clin. Lab Invest. (Suppl. 47) 21:770.Google Scholar
  3. Briggs, R., and King, T. J., 1960, Nucleocytoplasmic interactions in eggs and embryos, in: The Cell, Volume 1 (J. Brackett and A. E. Mirsky, eds.), Academic Press, New York, pp. 537–617.Google Scholar
  4. Brown, R. L., Wible, L. J., and Brinkley, B. R., 1980, Cytoplasmic microtubule assemblydisassembly in enucleated cells and regenerating karyoplasts, Cell Biol. Internatl. Rep. 4:453–458.CrossRefGoogle Scholar
  5. Bruno, J., Reich, N. R., and Lucas, J. J., 1981, Synthesis of globin in hybrid cells constructed by transplantation of dormant avian erythrocyte nuclei into enucleated fibroblasts, Mol. Cell Biol. 1:1163–1176.PubMedGoogle Scholar
  6. Carter, S. B., 1967, Effects of cytochalasins on mammalian cells, Nature 213:261–264.PubMedCrossRefGoogle Scholar
  7. Croce, C. M., and Koprowski, H., 1973, Enucleation of cells made simple and rescue of SV-40 by enucleated cells made even simpler, Virology 51:227–229.PubMedCrossRefGoogle Scholar
  8. deTerra, N., 1967, Macronuclear DNA synthesis in Stentor: Regulation by a cytoplasmic initiator, Proc. Natl. Acad. Sci. USA 57:607–614.CrossRefGoogle Scholar
  9. Ege, T., Hamberg, H., Krondahl, U., Ericson, J., and Ringertz, N. R., 1974, Characterization of minicells (nuclei) obtained by cytochalasin enucleation, Exp. Cell Res. 87:365–377.PubMedCrossRefGoogle Scholar
  10. Follett, E. A. C., 1974, A convenient method for enucleating cells in quantity, Exp. Cell Res. 84:72–78.PubMedCrossRefGoogle Scholar
  11. Follett, E. A. C., Pringle, C. R., Wunner, W. H., and Skehel, J. J., 1974, Virus replication in enucleate cells: Vesicular stomatitis virus and influenza virus, J. Virol. 13:394–399.PubMedGoogle Scholar
  12. Goldman, R. D., Pollack, R., and Hopkins, N. H., 1973, Preservation of normal behavior by enucleated cells in culture, Proc. Natl. Acad. Sci. USA 70:750–754.PubMedCrossRefGoogle Scholar
  13. Gurdon, J. B., 1962, Adult frogs derived from the nuclei of single somatic cells, Develop. Biol. 4:256–273.PubMedCrossRefGoogle Scholar
  14. Hightower, M. J., and Lucas, J. J., 1980, Construction of viable mouse-human hybrid cells by nuclear transplantation, J. Cell. Physiol. 105:93–103.PubMedCrossRefGoogle Scholar
  15. Hightower, M. J., Fairfield, E., and Lucas, J. J., 1981, A staining procedure for identifying viable cell hybrids constructed by somatic cell fusion, cybridization or nuclear transplantation, Somat. Cell Genet. 7:321–329.PubMedCrossRefGoogle Scholar
  16. Johnson, L. V, Walsh, M. L., and Chen, L. B., 1980, Localization of mitochondria in living cells with rhodamine 123, Proc. Natl. Acad. Sci. USA 77:990–994.PubMedCrossRefGoogle Scholar
  17. Lipsich, L. A., Lucas, J. J., and Kates, J. R., 1978, Cell cycle dependence of the reactivation of chick erythrocyte nuclei after transplantation into mouse L929 cell cytoplasts, J. Cell. Physiol. 97:199–208.PubMedCrossRefGoogle Scholar
  18. Lipsich, L. A., Lucas, J. J., and Kates, J. R., 1979a, Separation of cytoplasts and whole cells using density gradients of renografin, J. Cell. Physiol. 98:637–642.PubMedCrossRefGoogle Scholar
  19. Lipsich, L. A., Kates, J. R., and Lucas, J. J., 1979b, Expression of a liver-specific function by mouse fibroblast nuclei transplanted into rat hepatoma cytoplasts, Nature 281:74–76.PubMedCrossRefGoogle Scholar
  20. Lucas, J. J., 1982, Somatic cell hybridization, in: Eukaryotic Genes: Their Structure, Activity and Regulation (N. Maclean, S. P. Gregory, and R. A. Flavell, eds.), Butterworth, London.Google Scholar
  21. Lucas, J. J., and Kates, J. R., 1976, The construction of viable nuclear-cytoplasmic hybrid cells by nuclear transplantation, Cell 7:397–405.PubMedCrossRefGoogle Scholar
  22. Lucas, J. J., and Kates, J. R., 1977, Nuclear transplantation with mammalian cells, in: Methods in Cell Biology, Volume XV (D. M. Prescott, ed.), Academic Press, New York, pp. 359–370.Google Scholar
  23. Lucas, J. J., Szekely, E., and Kates, J. R., 1976, The regeneration and division of mouse L-cell karyoplasts, Cell 7:115–122.PubMedCrossRefGoogle Scholar
  24. Moser, G., Dorman, B. P., and Ruddle, F. H., 1975, Mouse-human heterokaryon analysis with a 33258 Hoechst-Giemsa technique, J. Cell Biol. 66:376–405.CrossRefGoogle Scholar
  25. Okada, M., Kleinman, A., and Schneiderman, H. A., 1974, Chimeric Drosophila adults produced by transplantation of nuclei into specific regions of fertilized eggs, Develop. Biol. 39:286–294.PubMedGoogle Scholar
  26. Peterson, J. A., and Weiss, M. C., 1972, Expression of differentiated functions in hepatoma cell hybrids: Induction of mouse albumin production in rat hepatoma-mouse fibroblast hybrids, Proc. Natl. Acad. Sci. USA 69:571–575.PubMedCrossRefGoogle Scholar
  27. Pollack, R., and Goldman, R., 1973, Synthesis of infective poliovirus in BSC-1 monkey cells enucleated with cytochalasin B, Science 179:915–916.PubMedCrossRefGoogle Scholar
  28. Prescott, D. M., Kates, J., and Kirkpatrick, J. B., 1971, Replication of vaccinia virus DNA in enucleated L-cells, J. Mol. Biol. 59:505–508.PubMedCrossRefGoogle Scholar
  29. Prescott, D. M., Myerson, D., and Wallace, J., 1972, Enucleation of mammalian cells with cytochalasin B, Exp. Cell Res. 71:480–485.PubMedCrossRefGoogle Scholar
  30. Ringertz, N. R., and Savage, R. E., 1976, Cell Hybrids, Academic Press, New York.Google Scholar
  31. Shay, J. W., Porter, K. R., and Prescott, D. M., 1974, The surface morphology and fine structure of CHO (Chinese hamster ovary) cells following enucleation, Proc. Natl. Acad. Sci. USA 71:3059–3063.PubMedCrossRefGoogle Scholar
  32. Veomett, G., Prescott, D. M., Shay, J., and Porter, K. R., 1974, Reconstruction of mammalian cells from nuclear and cytoplasmic components separated by treatment with cytochalasin B, Proc. Natl. Acad. Sci. USA 71:1999–2002.PubMedCrossRefGoogle Scholar
  33. Watkins, J. F., 1971, Fusion of cells for virus studies and production of cell hybrids, in: Methods in Virology, Volume 5 (T. K. Maramorasch and H. Koprowski, eds.), Academic Press, New York, pp. 1–32.Google Scholar
  34. Wessels, N. K., Spooner, B. S., Ash, J. F., Bradley, M. O., Luduena, M. A., Wrenn, E. L., and Yamada, K. M., 1971, Microfilaments in cellular and developmental processes: Contractile microfilament machinery of many cell types is reversibly inhibited by cytochalasin B, Science 171:135–143.CrossRefGoogle Scholar
  35. Wiktor, T. J., and Koprowski, H., 1974, Rhabdovirus replication in enucleated host cells, J. Virol. 14:300–306.PubMedGoogle Scholar
  36. Wright, W. E., and Hayflick, L., 1972, Formation of anucleate and multinucleate cells in normal and SV-40 transformed WI-38 by cytochalasin B, Exp. Cell Res. 74:187–194.PubMedCrossRefGoogle Scholar
  37. Zorn, G. A., and Anderson, C. W., 1981, Adenovirus type 2 expresses fiber in monkey-human hybrids and reconstructed cells, J. Virol. 37:759–769.PubMedGoogle Scholar
  38. Zorn, G. A., Lucas, J. J., and Kates, J. R., 1980, Purification and characterization of regnerating mouse L929 karyoplasts, Cell 18:659–672.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • Margaret J. Hightower
    • 1
  • Joseph J. Lucas
    • 1
  1. 1.Department of MicrobiologyState University of New York at Stony BrookStony BrookUSA

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