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Reprogramming of Somatic Cells Towards Pluripotency by Cell Fusion

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Polycomb Group Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1480))

Abstract

Pluripotent reprogramming can be dominantly induced in a somatic nucleus upon fusion with a pluripotent cell such as embryonic stem (ES) cell. Cell fusion between ES cells and somatic cells results in the formation of heterokaryons, in which the somatic nuclei begin to acquire features of the pluripotent partner. The generation of interspecies heterokaryons between mouse ES- and human somatic cells allows an experimenter to distinguish the nuclear events occurring specifically within the reprogrammed nucleus. Therefore, cell fusion provides a simple and rapid approach to look at the early nuclear events underlying pluripotent reprogramming. Here, we describe a polyethylene glycol (PEG)-mediated cell fusion protocol to generate interspecies heterokaryons and intraspecies hybrids between ES cells and B lymphocytes or fibroblasts.

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References

  1. Weiss MC, Green H (1967) Human-mouse hybrid cell lines containing partial complements of human chromosomes and functioning human genes. Proc Natl Acad Sci U S A 58(3):1104–1111

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256(5517):495–497

    Article  CAS  PubMed  Google Scholar 

  3. Tada M et al (2001) Nuclear reprogramming of somatic cells by in vitro hybridization with ES cells. Curr Biol 11(19):1553–1558

    Article  CAS  PubMed  Google Scholar 

  4. Cowan CA et al (2005) Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells. Science 309(5739):1369–1373

    Article  CAS  PubMed  Google Scholar 

  5. Piccolo FM et al (2011) Using heterokaryons to understand pluripotency and reprogramming. Philos Trans R Soc Lond B Biol Sci 366(1575):2260–2265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Soza-Ried J, Fisher AG (2012) Reprogramming somatic cells towards pluripotency by cellular fusion. Curr Opin Genet Dev 22(5):459–465

    Article  CAS  PubMed  Google Scholar 

  7. Pereira CF et al (2008) Heterokaryon-based reprogramming of human B lymphocytes for pluripotency requires Oct4 but not Sox2. PLoS Genet 4(9):e1000170

    Article  PubMed  PubMed Central  Google Scholar 

  8. Yamanaka S, Blau HM (2010) Nuclear reprogramming to a pluripotent state by three approaches. Nature 465(7299):704–712

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Terranova R et al (2006) Acquisition and extinction of gene expression programs are separable events in heterokaryon reprogramming. J Cell Sci 119(Pt 10):2065–2072

    Article  CAS  PubMed  Google Scholar 

  10. Pereira CF, Fisher AG (2009) Heterokaryon-based reprogramming for pluripotency. Curr Protoc Stem Cell Biol Chapter 4:Unit 4B.1

    PubMed  Google Scholar 

  11. Pereira CF et al (2010) ESCs require PRC2 to direct the successful reprogramming of differentiated cells toward pluripotency. Cell Stem Cell 6(6):547–556

    Article  CAS  PubMed  Google Scholar 

  12. Hui SW, Stenger DA (1993) Electrofusion of cells: hybridoma production by electrofusion and polyethylene glycol. Methods Enzymol 220:212–227

    Article  CAS  PubMed  Google Scholar 

  13. O’Connor MD et al (2008) Alkaline phosphatase-positive colony formation is a sensitive, specific, and quantitative indicator of undifferentiated human embryonic stem cells. Stem Cells 26(5):1109–1116

    Article  PubMed  Google Scholar 

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Correspondence to Amanda G. Fisher .

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© 2016 Springer Science+Business Media New York

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Malinowski, A.R., Fisher, A.G. (2016). Reprogramming of Somatic Cells Towards Pluripotency by Cell Fusion. In: Lanzuolo, C., Bodega, B. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 1480. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6380-5_25

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  • DOI: https://doi.org/10.1007/978-1-4939-6380-5_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6378-2

  • Online ISBN: 978-1-4939-6380-5

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