An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells

Nat Biotechnol. 2011 Aug 14;29(9):829-34. doi: 10.1038/nbt.1947.

Abstract

An important risk in the clinical application of human pluripotent stem cells (hPSCs), including human embryonic and induced pluripotent stem cells (hESCs and hiPSCs), is teratoma formation by residual undifferentiated cells. We raised a monoclonal antibody against hESCs, designated anti-stage-specific embryonic antigen (SSEA)-5, which binds a previously unidentified antigen highly and specifically expressed on hPSCs--the H type-1 glycan. Separation based on SSEA-5 expression through fluorescence-activated cell sorting (FACS) greatly reduced teratoma-formation potential of heterogeneously differentiated cultures. To ensure complete removal of teratoma-forming cells, we identified additional pluripotency surface markers (PSMs) exhibiting a large dynamic expression range during differentiation: CD9, CD30, CD50, CD90 and CD200. Immunohistochemistry studies of human fetal tissues and bioinformatics analysis of a microarray database revealed that concurrent expression of these markers is both common and specific to hPSCs. Immunodepletion with antibodies against SSEA-5 and two additional PSMs completely removed teratoma-formation potential from incompletely differentiated hESC cultures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / metabolism*
  • Biomarkers
  • Cell Differentiation
  • Cells, Cultured
  • Computational Biology
  • Flow Cytometry
  • Gene Expression Regulation
  • Humans
  • Immunohistochemistry
  • Mice
  • Mice, Inbred BALB C
  • Microarray Analysis
  • Pluripotent Stem Cells / chemistry
  • Pluripotent Stem Cells / cytology*
  • Polysaccharides / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Stage-Specific Embryonic Antigens / metabolism*
  • Teratoma / metabolism
  • Teratoma / pathology*

Substances

  • Antibodies, Monoclonal
  • Biomarkers
  • Polysaccharides
  • Stage-Specific Embryonic Antigens