Prospective isolation and characterization of mesenchymal stem cells from human placenta using a frizzled-9-specific monoclonal antibody.
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.
Battula
Venkata Lokesh
VL
Division of Hematology, Immunology, Oncology and Rheumatology, Department of Internal Medicine II, University Clinic of Tübingen, Tübingen, Germany.
Treml
Sabrina
S
Abele
Harald
H
Bühring
Hans-Jörg
HJ
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
D000911:Antibodies, Monoclonal; D017931:DNA Primers; C105574:FZD3 protein, human; D051157:Frizzled Receptors; D043562:Receptors, G-Protein-Coupled