Prospective isolation and characterization of mesenchymal stem cells from human placenta using a frizzled-9-specific monoclonal antibody

Differentiation. 2008 Apr;76(4):326-36. doi: 10.1111/j.1432-0436.2007.00225.x. Epub 2007 Oct 9.

Abstract

We have recently shown that frizzled-9 (FZD9, CD349) is expressed on the cell surface of cultured mesenchymal stromal cells (MSC) derived from the human bone marrow (BM) and chorionic placenta (PL). To study whether FZD9 is also a marker for naive mesenchymal stem cells (MSC), we analyzed the expression pattern of FZD9 on freshly isolated PL cells and determined the clonogenic potential of isolated FZD9(+) cells using the colony-forming units-fibroblastic (CFU-F) assay. About 0.2% of isolated PL cells were positive for FZD9. Two-color analysis revealed that FZD9(+) PL cells uniformly express CD9, CD63, and CD90, but are heterogeneous for CD10, CD13, and CD26 expression. In contrast to BM-derived MSC, PL-derived MSC expressed only low levels of CD271. Colony assays of sorted cells showed that clonogenic CFU-F reside exclusively in the FZD9(+) but not in the FZD9(-) fraction. Further analysis revealed that CFU-F were enriched by 60-fold in the FZD9(+)CD10(+)CD26(+) fraction but were absent in the FZD9(+)CD10(-)CD26(-) population. Cultured FZD9(+) cells expressed the embryonic stem cell makers Oct-4 and nanog as well as SSEA-4 and TRA1-2-49/6E. In addition, they could be differentiated into functional adipocytes and osteoblasts. This report describes for the first time that FZD9 is a novel and specific marker for the prospective isolation of MSC from human term PL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology
  • Antibodies, Monoclonal / immunology*
  • Base Sequence
  • Cell Differentiation
  • Chorion / cytology
  • DNA Primers
  • Female
  • Flow Cytometry
  • Frizzled Receptors / immunology*
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Osteoblasts / cytology
  • Placenta / cytology*
  • Receptors, G-Protein-Coupled / immunology*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Antibodies, Monoclonal
  • DNA Primers
  • FZD3 protein, human
  • Frizzled Receptors
  • Receptors, G-Protein-Coupled