Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis
Abstract Background The pharmaceutical agent pentosan polysulfate (PPS) is known to induce proliferation and chondrogenesis of mesenchymal progenitor cells (MPCs) in vitro and in vivo. However, the mechanism(s) of action of PPS in mediating these effects remains unresolved. In the present report we...
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| Format: | Article |
| Language: | English |
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BMC
2017-12-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-017-0723-y |
| _version_ | 1818422032900030464 |
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| author | Jiehua Wu Susan Shimmon Sharon Paton Christopher Daly Tony Goldschlager Stan Gronthos Andrew C. W. Zannettino Peter Ghosh |
| author_facet | Jiehua Wu Susan Shimmon Sharon Paton Christopher Daly Tony Goldschlager Stan Gronthos Andrew C. W. Zannettino Peter Ghosh |
| author_sort | Jiehua Wu |
| collection | DOAJ |
| description | Abstract Background The pharmaceutical agent pentosan polysulfate (PPS) is known to induce proliferation and chondrogenesis of mesenchymal progenitor cells (MPCs) in vitro and in vivo. However, the mechanism(s) of action of PPS in mediating these effects remains unresolved. In the present report we address this issue by investigating the binding and uptake of PPS by MPCs and monitoring gene expression and proteoglycan biosynthesis before and after the cells had been exposed to limited concentrations of PPS and then re-established in culture in the absence of the drug (MPC priming). Methods Immuno-selected STRO-1+ mesenchymal progenitor stem cells (MPCs) were prepared from human bone marrow aspirates and established in culture. The kinetics of uptake, shedding, and internalization of PPS by MPCs was determined by monitoring the concentration-dependent loss of PPS media concentrations using an enzyme-linked immunosorbent assay (ELISA) and the uptake of fluorescein isothiocyanate (FITC)-labelled PPS by MPCs. The proliferation of MPCs, following pre-incubation and removal of PPS (priming), was assessed using the Wst-8 assay method, and proteoglycan synthesis was determined by the incorporation of 35SO4 into their sulphated glycosaminoglycans. The changes in expression of MPC-related cell surface antigens of non-primed and PPS-primed MPCs from three donors was determined using flow cytometry. RNA sequencing of RNA isolated from non-primed and PPS-primed MPCs from the same donors was undertaken to identify the genes altered by the PPS priming protocol. Results The kinetic studies indicated that, in culture, PPS rapidly binds to MPC surface receptors, followed by internalisation and localization within the nucleus of the cells. Following PPS-priming of MPCs and a further 48 h of culture, both cell proliferation and proteoglycan synthesis were enhanced. Reduced expression of MPC-related cell surface antigen expression was promoted by the PPS priming, and RNA sequencing analysis revealed changes in the expression of 42 genes. Conclusion This study has shown that priming of MPCs with low concentrations of PPS enhanced chondrogenesis and MPC proliferation by modifying their characteristic basal gene and protein expression. These findings offer a novel approach to re-programming mesenchymal stem cells for clinical indications which require the repair or regeneration of cartilaginous tissues such as in osteoarthritis and degenerative disc disease. |
| first_indexed | 2024-12-14T13:19:49Z |
| format | Article |
| id | doaj.art-1788f54f9a194365987ae5d241964707 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-14T13:19:49Z |
| publishDate | 2017-12-01 |
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| series | Stem Cell Research & Therapy |
| spelling | doaj.art-1788f54f9a194365987ae5d2419647072022-12-21T22:59:58ZengBMCStem Cell Research & Therapy1757-65122017-12-018111510.1186/s13287-017-0723-yPentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesisJiehua Wu0Susan Shimmon1Sharon Paton2Christopher Daly3Tony Goldschlager4Stan Gronthos5Andrew C. W. Zannettino6Peter Ghosh7Proteobioactives Pty. Ltd.Proteobioactives Pty. Ltd.Myeloma Research Laboratory, Faculty of Health and Medical Sciences, University of Adelaide and the Cancer Theme, South Australia Health and Medical Research InstituteDepartment of Surgery, Monash UniversityDepartment of Surgery, Monash UniversityMesenchymal Stem Cell Laboratory, Adelaide Medical School, Faculty of Health and Medical Sciences, University of AdelaideMyeloma Research Laboratory, Faculty of Health and Medical Sciences, University of Adelaide and the Cancer Theme, South Australia Health and Medical Research InstituteProteobioactives Pty. Ltd.Abstract Background The pharmaceutical agent pentosan polysulfate (PPS) is known to induce proliferation and chondrogenesis of mesenchymal progenitor cells (MPCs) in vitro and in vivo. However, the mechanism(s) of action of PPS in mediating these effects remains unresolved. In the present report we address this issue by investigating the binding and uptake of PPS by MPCs and monitoring gene expression and proteoglycan biosynthesis before and after the cells had been exposed to limited concentrations of PPS and then re-established in culture in the absence of the drug (MPC priming). Methods Immuno-selected STRO-1+ mesenchymal progenitor stem cells (MPCs) were prepared from human bone marrow aspirates and established in culture. The kinetics of uptake, shedding, and internalization of PPS by MPCs was determined by monitoring the concentration-dependent loss of PPS media concentrations using an enzyme-linked immunosorbent assay (ELISA) and the uptake of fluorescein isothiocyanate (FITC)-labelled PPS by MPCs. The proliferation of MPCs, following pre-incubation and removal of PPS (priming), was assessed using the Wst-8 assay method, and proteoglycan synthesis was determined by the incorporation of 35SO4 into their sulphated glycosaminoglycans. The changes in expression of MPC-related cell surface antigens of non-primed and PPS-primed MPCs from three donors was determined using flow cytometry. RNA sequencing of RNA isolated from non-primed and PPS-primed MPCs from the same donors was undertaken to identify the genes altered by the PPS priming protocol. Results The kinetic studies indicated that, in culture, PPS rapidly binds to MPC surface receptors, followed by internalisation and localization within the nucleus of the cells. Following PPS-priming of MPCs and a further 48 h of culture, both cell proliferation and proteoglycan synthesis were enhanced. Reduced expression of MPC-related cell surface antigen expression was promoted by the PPS priming, and RNA sequencing analysis revealed changes in the expression of 42 genes. Conclusion This study has shown that priming of MPCs with low concentrations of PPS enhanced chondrogenesis and MPC proliferation by modifying their characteristic basal gene and protein expression. These findings offer a novel approach to re-programming mesenchymal stem cells for clinical indications which require the repair or regeneration of cartilaginous tissues such as in osteoarthritis and degenerative disc disease.http://link.springer.com/article/10.1186/s13287-017-0723-yMesenchymal progenitor cellsPentosan polysulfateHeparinChondrogenesisProliferationGene expression |
| spellingShingle | Jiehua Wu Susan Shimmon Sharon Paton Christopher Daly Tony Goldschlager Stan Gronthos Andrew C. W. Zannettino Peter Ghosh Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis Stem Cell Research & Therapy Mesenchymal progenitor cells Pentosan polysulfate Heparin Chondrogenesis Proliferation Gene expression |
| title | Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis |
| title_full | Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis |
| title_fullStr | Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis |
| title_full_unstemmed | Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis |
| title_short | Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis |
| title_sort | pentosan polysulfate binds to stro 1 mesenchymal progenitor cells is internalized and modifies gene expression a novel approach of pre programing stem cells for therapeutic application requiring their chondrogenesis |
| topic | Mesenchymal progenitor cells Pentosan polysulfate Heparin Chondrogenesis Proliferation Gene expression |
| url | http://link.springer.com/article/10.1186/s13287-017-0723-y |
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