Nuclear reprogramming of somatic nucleus hybridized with embryonic stem cells by electrofusion

Methods Mol Biol. 2006:329:411-20. doi: 10.1385/1-59745-037-5:411.

Abstract

Cell fusion is a powerful tool for understanding the molecular mechanisms of epigenetic reprogramming. In hybrid cells of somatic cells and pluripotential stem cells, including embryonic stem (ES) and embryonic germ cells, somatic nuclei acquire pluripotential competence. ES and embryonic germ cells retain intrinsic trans activity to induce epigenetic reprogramming. For generating hybrid cells, we have used the technique of electrofusion. Electrofusion is a highly effective, reproducible, and biomedically safe in vitro system. For successful cell fusion, two sequential steps of electric pulse stimulation are required for the alignment (pearl chain formation) of two different types of cells between electrodes in response to alternating current stimulation and for the fusion of cytoplasmic membranes by direct current stimulation. Optimal conditions for electrofusion with a pulse generator are introduced for ES and somatic cell fusion. Topics in the field of stem cell research include the successful production of cloned animals via the epigenetic reprogramming of somatic cells and contribution of spontaneous cell fusion to generating intrinsic plasticity of tissue stem cells. Cell fusion technology may make important contributions to the fields of epigenetic reprogramming and regenerative medicine.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cell Fusion / instrumentation
  • Cell Fusion / methods*
  • Cell Nucleus / genetics*
  • Clone Cells
  • Electricity
  • Embryo, Mammalian / cytology*
  • Epigenesis, Genetic*
  • Hybrid Cells / cytology
  • Hybrid Cells / metabolism*
  • Mice
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism*