Cytoplast containing reprogramming-related factors from human embryonic stem cells arrested at metaphase

Dev Growth Differ. 2011 Jan;53(1):18-25. doi: 10.1111/j.1440-169X.2010.01217.x.

Abstract

Generating pluripotent stem cells directly from a patient's somatic cells is one of the major methods to avoid rejection in future regenerative medicine. It is reported that human embryonic stem cells (hESCs) are able to reprogram the nuclei of fully differentiated human somatic cells, apparently conferring on them a pluripotent state. However, the ability of the cytoplasts from enucleated hESCs to reprogram somatic cells causes much controversy. Here we detect the location of pluripotency-related factors such as Oct4/Nanog/Sox2 in the hESCs at division and non-division stage and obtain the cytoplasts of hESCs by centrifugation. We demonstrate for the first time that the cytoplast from hESCs arrested at the division phase of cell the cycle contains the reprogramming factors and this kind of cytoplast can be obtained through gradient centrifugation. These give us direct proof of the possibility of reprogramming somatic cell using cytoplast of hESCs and make this a possible method for getting patient-specific pluripotent cells without extrinsic DNA introduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Cellular Reprogramming / physiology*
  • Cytoplasm / metabolism*
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism*
  • Flow Cytometry
  • Homeodomain Proteins / metabolism
  • Humans
  • Metaphase / physiology
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3 / metabolism
  • SOXB1 Transcription Factors / metabolism
  • Transcription Factors / metabolism*

Substances

  • Homeodomain Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Transcription Factors