title: Prospective isolation and characterization of mesenchymal stem cells from human placenta using a frizzled-9-specific monoclonal antibody.
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.
authors.affiliation: Division of Hematology, Immunology, Oncology and Rheumatology, Department of Internal Medicine II, University Clinic of Tübingen, Tübingen, Germany.
authors.forename: Venkata Lokesh
authors.initials: VL
authors.lastname: Battula
authors.forename: Sabrina
authors.initials: S
authors.lastname: Treml
authors.forename: Harald
authors.initials: H
authors.lastname: Abele
authors.forename: Hans-Jörg
authors.initials: HJ
authors.lastname: Bühring
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D002454:Cell Differentiation; D002823:Chorion / Q000166:cytology; D017931:DNA Primers; D005260:Female; D005434:Flow Cytometry; D051157:Frizzled Receptors / Q000276:immunology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology; D010920:Placenta / Q000166:cytology*; D043562:Receptors, G-Protein-Coupled / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; C105574:FZD3 protein, human; D051157:Frizzled Receptors; D043562:Receptors, G-Protein-Coupled