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Type: Journal article
Title: Heat shock protein-90 beta is expressed at the surface of multipotential mesenchymal precursor cells: Generation of a novel monoclonal antibody, STRO-4, with specificity for mesenchymal precursor cells from human and ovine tissues
Author: Gronthos, S.
McCarty, R.
Mrozik, K.
Fitter, S.
Paton, S.
Menicanin, D.
Itescu, S.
Bartold, P.
Xian, C.
Zannettino, A.
Citation: Stem Cells and Development, 2009; 18(9):1253-1261
Publisher: Mary Ann Liebert Inc Publ
Issue Date: 2009
ISSN: 1547-3287
Statement of
Stan Gronthos, Rosa McCarty, Krzysztof Mrozik, Stephen Fitter, Sharon Paton, Danijela Menicanin, Silviu Itescu, P. Mark Bartold, Cory Xian and Andrew C.W. Zannettino
Abstract: Mesenchymal stromal cells (MSCs) and their precursor cells (MPCs) can proliferate and differentiate into multiple mesodermal and some ectodermal and endodermal tissues. Culture-expanded MSCs are currently being evaluated as a possible cell therapy to replace/repair injured or diseased tissues. While a number of mAb reagents with specificity to human MSCs, including STRO-1, STRO-3 (BLK ALP), CD71 (SH2, SH3), CD106 (VCAM-1), CD166, and CD271, have facilitated the isolation of purified populations of human MSCs from primary tissues, few if any mAb reagents have been described that can be used to isolate equivalent cells from other species. This is of particular relevance when assessing the tissue regenerative efficacy of MSCs in large immunocompetent, preclinical animal models of disease. In light of this, we sought to generate novel monoclonal antibodies (mAb) with specific reactivity against a cell surface molecule that is expressed at high levels by MSCs from different species. Using CD106 (VCAM-1)-selected ovine MSCs as an immunogen, mAb-producing hybridomas were selected for their reactivity to both human and ovine MSCs. One such hybridoma, termed STRO-4, produced an IgG mAb that reacted with <5% of human and ovine bone marrow (BM) mononuclear cells. As a single selection reagent, STRO-4 mAb was able to enrich colony-forming fibroblasts (CFU-F) in both human and ovine BM by 16- and 8-folds, respectively. Cells isolated with STRO-4 exhibited reactivity with markers commonly associated with MSCs isolated by plastic adherence including CD29, CD44, and CD166. Moreover, when placed in inductive culture conditions in vitro, STRO-4+ MSCs exhibited multilineage differentiation potential and were capable of forming a mineralized matrix, lipid-filled adipocytes, and chondrocytes capable of forming a glycosaminoglycan-rich matrix. Biochemical analysis revealed that STRO-4 identified the beta isoform of heat shock protein-90 (Hsp90β). In addition to identifying an antibody reagent that identifies a highly conserved epitope expressed by MSCs from different species, our study also points to a potential role for Hsp90β in MSC biology.
Keywords: Leukocytes, Mononuclear
Bone Marrow Cells
Cell Membrane
Mesenchymal Stem Cells
Multipotent Stem Cells
Mice, Inbred BALB C
Antibodies, Monoclonal
Flow Cytometry
Cell Differentiation
Antibody Specificity
HSP90 Heat-Shock Proteins
Adult Stem Cells
DOI: 10.1089/scd.2008.0400
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