Prospective isolation and characterization of mesenchymal stem cells from human placenta using a frizzled-9-specific monoclonal antibody |
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Authors: | Battula Venkata Lokesh Treml Sabrina Abele Harald Bühring Hans-Jörg |
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Affiliation: | Division of Hematology, Immunology, Oncology and Rheumatology, Department of Internal Medicine II, University Clinic of Tübingen, Tübingen, Germany Tel: +49 7071 2982730 Fax: +49 7071 292730; Department of Obstetrics and Gynecology, University Clinic of Tübingen, Tübingen, Germany |
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Abstract: | 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. |
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Keywords: | placenta mesenchymal stem cells multiparameter flow cytometry frizzled-9 |
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