Cell Fusion pp 165-184 | Cite as

Analyzing Cell Fusion Events Within the Central Nervous System Using Bone Marrow Chimerism

Part of the Methods in Molecular Biology book series (MIMB, volume 1313)


It has emerged that cells which typically reside in the bone marrow have the capacity to cross the blood brain barrier and contribute genetic material to a range of neuronal cell types within the central nervous system. One such mechanism to account for this phenomenon is cellular fusion, occurring between migrating bone marrow-derived stem cells and neuronal cells in-situ. Biologically, the significance as to why cells from distinct lineages fuse with cells of the central nervous system is, as yet, unclear. Growing evidence however suggests that these cell fusion events could provide an efficient means of rescuing the highly complex and differentiated neuronal cell types that cannot be replaced in adulthood. To facilitate further understanding of cell fusion within the central nervous system, we describe here a technique to establish chimeric mice that are stably reconstituted with green fluorescent protein expressing sex-mismatched bone marrow. These chimeric mice are known to represent an excellent model for studying bone marrow cell migration and infiltration throughout the body, while in parallel, as will be described here, also provide a means to neatly analyze both bone marrow-derived cell fusion and trans-differentiation events within the central nervous system.

Key words

Bone marrow transplant Stem cells Central nervous system Neurons Purkinje cells Fusion Heterokaryon Synkaryon Green fluorescent protein Chimerism 



This work is supported by a project grants from the Medical Research Council and the University Research Council (University of Bristol).


  1. 1.
    Mezey E, Chandross KJ, Harta G et al (2000) Turning blood into brain: cells bearing neuronal antigens generated in vivo from bone marrow. Science 290:1779–1782CrossRefPubMedGoogle Scholar
  2. 2.
    Brazelton TR, Rossi FM, Keshet GI et al (2000) From marrow to brain: expression of neuronal phenotypes in adult mice. Science 290:1775–1779CrossRefPubMedGoogle Scholar
  3. 3.
    Priller J, Persons DA, Klett FF et al (2001) Neogenesis of cerebellar Purkinje neurons from gene-marked bone marrow cells in vivo. J Cell Biol 155:733–738CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Nakano K, Migita M, Mochizuki H et al (2001) Differentiation of transplanted bone marrow cells in the adult mouse brain. Transplantation 71:1735–1740CrossRefPubMedGoogle Scholar
  5. 5.
    Alvarez-Dolado M, Pardal R, Garcia-Verdugo JM et al (2003) Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature 425:968–973CrossRefPubMedGoogle Scholar
  6. 6.
    Weimann JM, Johansson CB, Trejo A et al (2003) Stable reprogrammed heterokaryons form spontaneously in Purkinje neurons after bone marrow transplant. Nat Cell Biol 5:959–966CrossRefPubMedGoogle Scholar
  7. 7.
    Wiersema A, Dijk F, Dontje B et al (2007) Cerebellar heterokaryon formation increases with age and after irradiation. Stem Cell Res 1:150–154CrossRefPubMedGoogle Scholar
  8. 8.
    Magrassi L, Grimaldi P, Ibatici A et al (2007) Induction and survival of binucleated Purkinje neurons by selective damage and aging. J Neurosci 27:9885–9892CrossRefPubMedGoogle Scholar
  9. 9.
    Johansson CB, Youssef S, Koleckar K et al (2008) Extensive fusion of haematopoietic cells with Purkinje neurons in response to chronic inflammation. Nat Cell Biol 10:575–583CrossRefPubMedCentralPubMedGoogle Scholar
  10. 10.
    Espejel S, Romero R, Alvarez-Buylla A (2009) Radiation damage increases Purkinje neuron heterokaryons in neonatal cerebellum. Ann Neurol 66:100–109CrossRefPubMedGoogle Scholar
  11. 11.
    Nern C, Wolff I, Macas J et al (2009) Fusion of hematopoietic cells with Purkinje neurons does not lead to stable heterokaryon formation under noninvasive conditions. J Neurosci 29:3799–3807CrossRefPubMedGoogle Scholar
  12. 12.
    Weimann JM, Charlton CA, Brazelton TR et al (2003) Contribution of transplanted bone marrow cells to Purkinje neurons in human adult brains. Proc Natl Acad Sci U S A 100:2088–2093CrossRefPubMedCentralPubMedGoogle Scholar
  13. 13.
    Zhu X, Siedlak SL, Wang Y et al (2008) Neuronal binucleation in Alzheimer disease hippocampus. Neuropathol Appl Neurobiol 34:457–465CrossRefPubMedGoogle Scholar
  14. 14.
    Kemp K, Gray E, Wilkins A et al (2012) Purkinje cell fusion and binucleate heterokaryon formation in multiple sclerosis cerebellum. Brain 135:2962–2972CrossRefPubMedGoogle Scholar
  15. 15.
    Chen KA, Cruz PE, Lanuto DJ et al (2011) Cellular fusion for gene delivery to SCA1 affected Purkinje neurons. Mol Cell Neurosci 47:61–70CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    Corti S, Locatelli F, Donadoni C et al (2004) Wild-type bone marrow cells ameliorate the phenotype of SOD1-G93A ALS mice and contribute to CNS, heart and skeletal muscle tissues. Brain 127:2518–2532CrossRefPubMedGoogle Scholar
  17. 17.
    Duran-Struuck R, Dysko RC (2009) Principles of bone marrow transplantation (BMT): providing optimal veterinary and husbandry care to irradiated mice in BMT studies. J Am Assoc Lab Anim Sci 48:11–22PubMedCentralPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Multiple Sclerosis and Stem Cell Group, School of Clinical SciencesUniversity of BristolBristolUK

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