Secreted cerberus1 as a marker for quantification of definitive endoderm differentiation of the pluripotent stem cells

PLoS One. 2013 May 22;8(5):e64291. doi: 10.1371/journal.pone.0064291. Print 2013.

Abstract

To date, CXCR4 and E-cadherin double-positive cells detected by flow cytometry have been used to identify the differentiation of embryonic stem (ES) cells or induced pluripotent stem (iPS) cells into definitive endoderm (DE) lineages. Quantification of DE differentiation from ES/iPS cells by using flow cytometry is a multi-step procedure including dissociation of the cells, antibody reaction, and flow cytometry analysis. To establish a quick assay method for quantification of ES/iPS cell differentiation into the DE without dissociating the cells, we examined whether secreted Cerberus1 (Cer1) protein could be used as a marker. Cer1 is a secreted protein expressed first in the anterior visceral endoderm and then in the DE. The amount of Cer1 secreted correlated with the proportion of CXCR4+/E-Cadherin+ cells that differentiated from mouse ES cells. In addition, we found that human iPS cell-derived DE also expressed the secreted CER1 and that the expression level correlated with the proportion of SOX17+/FOXA2+ cells present. Taken together, these results show that Cer1 (or CER1) serves as a good marker for quantification of DE differentiation of mouse and human ES/iPS cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Biomarkers / metabolism*
  • Cell Differentiation*
  • Cell Line
  • Cytokines / metabolism*
  • DNA Primers
  • Endoderm / cytology*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Mice
  • Pluripotent Stem Cells / cytology*
  • Proteins / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Biomarkers
  • CER1 protein, human
  • Cer1 protein, mouse
  • Cytokines
  • DNA Primers
  • Proteins

Grants and funding

This work was supported by a Grant-in-Aid (#21390280 to SK and NEXT program from JST to SK), National Institute of Biomedical Innovation (to NS), and, in part, by a Global COE grant (Cell Fate Regulation Research and Education Unit). SK received funding from the Dojindo Laboratories. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.