Transfer of Macromolecules Using Erythrocyte Ghosts

  • Martin C. Rechsteiner


Microinjection using RBC ghosts is accomplished by first introducing molecules into RBCs during hypotonie hemolysis and then fusing the RBCs to cultured cells with Sendai virus or polyethylene glycol. In this manner molecules can be injected into large numbers of cultured cells within a short time. The method was developed several years ago in three laboratories (Furusawa et al., 1974; Schlegel and Rechsteiner, 1975; Loyter et al., 1975), and it has been used to study a variety of cellular, genetic, and biochemical problems. Since anyone reading this chapter probably has an application in mind, the text emphasizes methodology. Several articles are available which review previous applications and present a broader discussion of the technique (Kulka and Loyter, 1979; Furusawa, 1980). Those who wish to use RBC-mediated injection should consider microneedles or liposomes as alternate injection procedures. Each technique offers certain advantages and disadvantages, and these are briefly reviewed.


Hank Balance Salt Solution Sendai Virus Erythrocyte Ghost Cell BioI Hypotonic Saline 
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  1. Anderson, W. F., Deisseroth, A., Nienhuis, A. W., Gopalakrishnan, T. V., Huang, A., and Krueger, L., 1978, Cellular and molecular studies on globin gene expression, Natl. Cancer Inst. Monogr. 48:65.PubMedGoogle Scholar
  2. Capecchi, M. R., Von der Haar, R. A., Capecchi, N. E., and Sveda, M. M., 1977, The isolation of a suppressible nonsense mutant in mammalian cells, Cell 12:371.PubMedCrossRefGoogle Scholar
  3. Eickbush, T. H., and Moudrianakis, E. N., 1978, The compaction of DNA helices into either continuous supercoils or folded fiber rods and torroids, Cell 13:295.PubMedCrossRefGoogle Scholar
  4. Fischer, T. M., Hoest, C. W. M., Stohr-Liesen, M., Schmid-Schonbein, H., and Skalak, R., 1981, The stress-free shape of the red blood cell membrane, Biophys. J. 34:409.PubMedCrossRefGoogle Scholar
  5. Furusawa, M., 1980, Cellular microinjection by cell fusion: Technique and application in biology and medicine, Int. Rev. Cytol. 62:29.PubMedCrossRefGoogle Scholar
  6. Furusawa, M., Nishimura, T., Yamaizumi, M., and Okada, Y., 1974, Injection of foreign substances into single cells by cell fusion, Nature 249:449.PubMedCrossRefGoogle Scholar
  7. Harris, H., and Watkins, J. F., 1965, Hybrid cells derived from mouse and man: Artificial heterokaryons of mammalian cells from different species, Nature 205:640–646.PubMedCrossRefGoogle Scholar
  8. Ihler, G., Glew, R., and Schnure, F., 1973, Enzyme loading of erythrocytes, Proc. Natl. Acad. Sci. USA 70:2663.PubMedCrossRefGoogle Scholar
  9. Kaltoft, K., and Celis, J. E., 1978, Ghost-mediated transfer of human hypoxanthine-quanine phosphoribosyl transferase into deficient Chinese hamster ovary cells by means of polyethylene glycol-induced fusion, Exp. Cell Res. 115:423.PubMedCrossRefGoogle Scholar
  10. Kaltoft, K., Zeuthen, J., Engback, F, Piper, P. W., and Celis, J. E., 1976, Transfer of tRNAs to somatic cells mediated by Sendai virus induced fusion, Proc. Natl. Acad. Sci. USA 73:2793.PubMedCrossRefGoogle Scholar
  11. Kriegler, M. P., and Livingston, D. M., 1977, Chemically facilitated microinjection of proteins into intact monolayers of tissue culture cells, Somat. Cell Genet. 3:603.PubMedCrossRefGoogle Scholar
  12. Kulka, R. G., and Loyter, A., 1979, The use of fusion methods for the microinjection of animal cells, in: Current Topics in Membranes and Transport, Volume 12 (F. Bronner and A. Kleinzeller, eds.), Academic Press, New York, p. 365.Google Scholar
  13. Laemmli, U. K., 1975, Characterization of DNA condensates induced by poly (ethylene oxide) and polylysine, Proc. Natl. Acad. Sci. USA 72:4288.PubMedCrossRefGoogle Scholar
  14. LoBue, J., Dornfest, B. S., Gordon, A. S., Hurst, J., and Quastler, H., 1963, Marrow distribution in rat femurs determined in cell enumeration and Fe59 labeling, Proc. Soc. Exp. Biol. Med. 112:1058.PubMedGoogle Scholar
  15. Loyter, A., Zakai, N., and Kulka, R., 1975, “Ultramicroinjection” of macromolecules or small particles into animal cells, J. Cell Biol. 66:292.PubMedCrossRefGoogle Scholar
  16. Maniatis, T., Venable, Jr., J. H., and Lerman, L. S., 1974, The structure of Ψ DNA, J. Mol. Biol. 84:37.PubMedCrossRefGoogle Scholar
  17. Mercer, W. E., Terefinko, D. J., and Schlegel, R. A., 1979, Red cell-mediated microinjection of macromolecules into monolayer cultures of mammalian cells, Cell Biol. Internatl. Rep. 3:265.CrossRefGoogle Scholar
  18. Peretz, H., Toister, Z., Laster, Y., and Loyter, A., 1974, Fusion of intact human erythrocytes and erythrocyte ghosts, J. Cell Biol. 63:1.PubMedCrossRefGoogle Scholar
  19. Rechsteiner, M., 1975, Uptake of proteins by red blood cells, Exp. Cell Res. 93:487.PubMedCrossRefGoogle Scholar
  20. Schlegel, R., and Rechsteiner, M., 1975, Microinjection of thymidine kinase and bovine serum albumin into mammalian cells by fusion with red blood cells. Cell 3:371.CrossRefGoogle Scholar
  21. Schlegel, R. A., Iverson, P., and Rechsteiner, M. C., 1978, The turnover of tRNAs microinjected into animal cells, Nucleic Acids Res. 5:3715.PubMedCrossRefGoogle Scholar
  22. Seeman, P., 1967, Transient holes in the erythrocyte membrane during hypotonie hemolysis and stable holes in the membrane after lysis by saponin and lysolecithin, J. Cell Biol. 32:55.PubMedCrossRefGoogle Scholar
  23. Steinman, R. M., Silver, J. M., and Cohn, Z. A., 1974, Pinocytosis in fibroblasts: Quantitative studies in vitro, J. Cell. Biol. 63:949.PubMedCrossRefGoogle Scholar
  24. Straus, S. E., and Raskas, H. J., 1980, Transfection of KB cells by polyethylene glycol-induced fusion with erythrocyte ghosts containing adenovirus type 2 DNA, J. Gen. Virol. 48:241.PubMedCrossRefGoogle Scholar
  25. Wasserman, M., Zakai, N., Loyter, A., and Kulka, R. G., 1976, A quantitative study of ultramicroinjection of macromolecules into animal cells, Cell 7:551.PubMedCrossRefGoogle Scholar
  26. Watkins, J. F., 1971, Fusion of cells for virus studies and production of cell hybrids, in: Methods in Virology, Volume V (K. Maramorosch and H. Koprowski, eds.), Academic Press, New York, p. 1.Google Scholar
  27. Yamaizumi, M., Uchida, T., Okada, Y., and Furusawa, M., 1978, Neutralization of diphtheria toxin in living cells by microinjection of antifragment A contained within resealed erythrocyte ghosts, Cell 13:227.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • Martin C. Rechsteiner
    • 1
  1. 1.Department of BiologyUniversity of UtahSalt Lake CityUSA

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