Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency
The capacity to produce therapeutically relevant quantities of multipotent mesenchymal stromal cells (MSCs) via in vitro culture is a common prerequisite for stem cell-based therapies. Although culture expanded MSCs are widely studied and considered for therapeutic applications, it has remained chal...
| Main Authors: | , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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National Academy of Sciences (U.S.)
2015
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| Online Access: | http://hdl.handle.net/1721.1/96533 https://orcid.org/0000-0001-5735-0560 https://orcid.org/0000-0001-7215-1439 |
| _version_ | 1826191964173762560 |
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| author | Lee, Wong Cheng J. Shi, Hui Poon, Zhiyong Nyan, Lin Myint Kaushik, Tanwi Shivashankar, G. V. Chan, Jerry K. Y. Lim, Chwee Teck Han, Jongyoon Van Vliet, Krystyn J |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Lee, Wong Cheng J. Shi, Hui Poon, Zhiyong Nyan, Lin Myint Kaushik, Tanwi Shivashankar, G. V. Chan, Jerry K. Y. Lim, Chwee Teck Han, Jongyoon Van Vliet, Krystyn J |
| author_sort | Lee, Wong Cheng J. |
| collection | MIT |
| description | The capacity to produce therapeutically relevant quantities of multipotent mesenchymal stromal cells (MSCs) via in vitro culture is a common prerequisite for stem cell-based therapies. Although culture expanded MSCs are widely studied and considered for therapeutic applications, it has remained challenging to identify a unique set of characteristics that enables robust identification and isolation of the multipotent stem cells. New means to describe and separate this rare cell type and its downstream progenitor cells within heterogeneous cell populations will contribute significantly to basic biological understanding and can potentially improve efficacy of stem and progenitor cell-based therapies. Here, we use multivariate biophysical analysis of culture-expanded, bone marrow-derived MSCs, correlating these quantitative measures with biomolecular markers and in vitro and in vivo functionality. We find that, although no single biophysical property robustly predicts stem cell multipotency, there exists a unique and minimal set of three biophysical markers that together are predictive of multipotent subpopulations, in vitro and in vivo. Subpopulations of culture-expanded stromal cells from both adult and fetal bone marrow that exhibit sufficiently small cell diameter, low cell stiffness, and high nuclear membrane fluctuations are highly clonogenic and also exhibit gene, protein, and functional signatures of multipotency. Further, we show that high-throughput inertial microfluidics enables efficient sorting of committed osteoprogenitor cells, as distinct from these mesenchymal stem cells, in adult bone marrow. Together, these results demonstrate novel methods and markers of stemness that facilitate physical isolation, study, and therapeutic use of culture-expanded, stromal cell subpopulations. |
| first_indexed | 2024-09-23T09:04:04Z |
| format | Article |
| id | mit-1721.1/96533 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:04:04Z |
| publishDate | 2015 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/965332022-09-26T10:13:00Z Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency Lee, Wong Cheng J. Shi, Hui Poon, Zhiyong Nyan, Lin Myint Kaushik, Tanwi Shivashankar, G. V. Chan, Jerry K. Y. Lim, Chwee Teck Han, Jongyoon Van Vliet, Krystyn J Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Department of Materials Science and Engineering Singapore-MIT Alliance in Research and Technology (SMART) Han, Jongyoon Van Vliet, Krystyn J. The capacity to produce therapeutically relevant quantities of multipotent mesenchymal stromal cells (MSCs) via in vitro culture is a common prerequisite for stem cell-based therapies. Although culture expanded MSCs are widely studied and considered for therapeutic applications, it has remained challenging to identify a unique set of characteristics that enables robust identification and isolation of the multipotent stem cells. New means to describe and separate this rare cell type and its downstream progenitor cells within heterogeneous cell populations will contribute significantly to basic biological understanding and can potentially improve efficacy of stem and progenitor cell-based therapies. Here, we use multivariate biophysical analysis of culture-expanded, bone marrow-derived MSCs, correlating these quantitative measures with biomolecular markers and in vitro and in vivo functionality. We find that, although no single biophysical property robustly predicts stem cell multipotency, there exists a unique and minimal set of three biophysical markers that together are predictive of multipotent subpopulations, in vitro and in vivo. Subpopulations of culture-expanded stromal cells from both adult and fetal bone marrow that exhibit sufficiently small cell diameter, low cell stiffness, and high nuclear membrane fluctuations are highly clonogenic and also exhibit gene, protein, and functional signatures of multipotency. Further, we show that high-throughput inertial microfluidics enables efficient sorting of committed osteoprogenitor cells, as distinct from these mesenchymal stem cells, in adult bone marrow. Together, these results demonstrate novel methods and markers of stemness that facilitate physical isolation, study, and therapeutic use of culture-expanded, stromal cell subpopulations. National University of Singapore (Graduate School for Integrative Sciences and Engineering Program) Singapore-MIT Alliance (Singapore-MIT Alliance-3 graduate fellowship program) Singapore. National Research Foundation Singapore-MIT Alliance for Research and Technology (BioSystems and Micromechanics Interdisciplinary Research Group) Singapore. National Medical Research Council (NMRC/Clinician Scientist Award/012/2009) 2015-04-13T16:40:31Z 2015-04-13T16:40:31Z 2014-10 2014-02 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/96533 Lee, Wong Cheng, Hui Shi, Zhiyong Poon, Lin Myint Nyan, Tanwi Kaushik, G. V. Shivashankar, Jerry K. Y. Chan, Chwee Teck Lim, Jongyoon Han, and Krystyn J. Van Vliet. “Multivariate Biophysical Markers Predictive of Mesenchymal Stromal Cell Multipotency.” Proceedings of the National Academy of Sciences 111, no. 42 (October 8, 2014): E4409–E4418. 25298531 https://orcid.org/0000-0001-5735-0560 https://orcid.org/0000-0001-7215-1439 en_US http://dx.doi.org/10.1073/pnas.1402306111 Proceedings of the National Academy of Sciences Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences (U.S.) National Academy of Sciences (U.S.) |
| spellingShingle | Lee, Wong Cheng J. Shi, Hui Poon, Zhiyong Nyan, Lin Myint Kaushik, Tanwi Shivashankar, G. V. Chan, Jerry K. Y. Lim, Chwee Teck Han, Jongyoon Van Vliet, Krystyn J Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| title | Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| title_full | Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| title_fullStr | Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| title_full_unstemmed | Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| title_short | Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| title_sort | multivariate biophysical markers predictive of mesenchymal stromal cell multipotency |
| url | http://hdl.handle.net/1721.1/96533 https://orcid.org/0000-0001-5735-0560 https://orcid.org/0000-0001-7215-1439 |
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