High-efficiency stem cell fusion-mediated assay reveals Sall4 as an enhancer of reprogramming

Connie C. Wong, Alexandre Gaspar Maia, Miguel Ramalho-Santos, Renee A. Reijo Pera

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Several methods allow reprogramming of differentiated somatic cells to embryonic stem cell-like cells. However, the process of reprogramming remains ineffecient and the underlying molecular mechanisms are poorly understood. Here, we report the optimisation of somatic cell fusion with embryonic stem cells in order to provide an efficient, quantitative assay to screen for factors that facilitate reprogramming. Following optimization, we achieved a reprogramming efficiency 15-590 fold higher than previous protocols. This allowed observation of cellular events during the reprogramming process. Moreover, we demonstate that overexpression of the Spalt transcription factor, Sall4, which was previously identified as a regulator of embryonic stem cell pluripotency and early mouse development, can enhance reprogramming. The reprogramming activity of Sall4 is independent of an N-terminal domain implicated in recruiting the nucleosome remodelling and deacetylase corepressor complex, a global transcriptional repressor. These results indicate that improvements in reprogramming assays including fusion assays, may allow the systematic identification and molecular characterization of enhancers of somatic cell reprogramming.

Original languageEnglish (US)
Article numbere1955
JournalPLoS One
Volume3
Issue number4
DOIs
StatePublished - Apr 16 2008
Externally publishedYes

Fingerprint

cell fusion
Cell Fusion
embryonic stem cells
Embryonic Stem Cells
Stem cells
somatic cells
stem cells
Assays
Fusion reactions
Stem Cells
Mi-2 Nucleosome Remodeling and Deacetylase Complex
assays
Co-Repressor Proteins
nucleosomes
Nucleosomes
Transcription Factors
transcription factors
Observation
mice
cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

High-efficiency stem cell fusion-mediated assay reveals Sall4 as an enhancer of reprogramming. / Wong, Connie C.; Gaspar Maia, Alexandre; Ramalho-Santos, Miguel; Reijo Pera, Renee A.

In: PLoS One, Vol. 3, No. 4, e1955, 16.04.2008.

Research output: Contribution to journalArticle

Wong, Connie C. ; Gaspar Maia, Alexandre ; Ramalho-Santos, Miguel ; Reijo Pera, Renee A. / High-efficiency stem cell fusion-mediated assay reveals Sall4 as an enhancer of reprogramming. In: PLoS One. 2008 ; Vol. 3, No. 4.
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