Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples
Mesenchymal stem cells (MSC) make up less than 1% of the bone marrow (BM). Several methods are used for their isolation such as gradient separation or centrifugation, but these methodologies are not direct and, thus, plastic adherence outgrowth or magnetic/fluorescent-activated sorting is required....
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MDPI AG
2022-01-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/9/2/49 |
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| author | Silvia Zia Carola Cavallo Ilaria Vigliotta Valentina Parisi Brunella Grigolo Roberto Buda Pasquale Marrazzo Francesco Alviano Laura Bonsi Andrea Zattoni Pierluigi Reschiglian Barbara Roda |
| author_facet | Silvia Zia Carola Cavallo Ilaria Vigliotta Valentina Parisi Brunella Grigolo Roberto Buda Pasquale Marrazzo Francesco Alviano Laura Bonsi Andrea Zattoni Pierluigi Reschiglian Barbara Roda |
| author_sort | Silvia Zia |
| collection | DOAJ |
| description | Mesenchymal stem cells (MSC) make up less than 1% of the bone marrow (BM). Several methods are used for their isolation such as gradient separation or centrifugation, but these methodologies are not direct and, thus, plastic adherence outgrowth or magnetic/fluorescent-activated sorting is required. To overcome this limitation, we investigated the use of a new separative technology to isolate MSCs from BM; it label-free separates cells based solely on their physical characteristics, preserving their native physical properties, and allows real-time visualization of cells. BM obtained from patients operated for osteochondral defects was directly concentrated in the operatory room and then analyzed using the new technology. Based on cell live-imaging and the sample profile, it was possible to highlight three fractions (F1, F2, F3), and the collected cells were evaluated in terms of their morphology, phenotype, CFU-F, and differentiation potential. Multipotent MSCs were found in F1: higher CFU-F activity and differentiation potential towards mesenchymal lineages compared to the other fractions. In addition, the technology depletes dead cells, removing unwanted red blood cells and non-progenitor stromal cells from the biological sample. This new technology provides an effective method to separate MSCs from fresh BM, maintaining their native characteristics and avoiding cell manipulation. This allows selective cell identification with a potential impact on regenerative medicine approaches in the orthopedic field and clinical applications. |
| first_indexed | 2024-03-09T22:36:19Z |
| format | Article |
| id | doaj.art-f83680c90c3e47e5b639fc469e7cd9ed |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-09T22:36:19Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-f83680c90c3e47e5b639fc469e7cd9ed2023-11-23T18:48:18ZengMDPI AGBioengineering2306-53542022-01-01924910.3390/bioengineering9020049Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow SamplesSilvia Zia0Carola Cavallo1Ilaria Vigliotta2Valentina Parisi3Brunella Grigolo4Roberto Buda5Pasquale Marrazzo6Francesco Alviano7Laura Bonsi8Andrea Zattoni9Pierluigi Reschiglian10Barbara Roda11Stem Sel Ltd., Viale Giuseppe Fanin 48, 40127 Bologna, ItalyLaboratory RAMSES, Research & Innovation Technology Department, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, ItalyUnit of Histology, Embryology and Applied Biology, Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via Belmeloro 8, 40126 Bologna, ItalyLaboratory RAMSES, Research & Innovation Technology Department, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, ItalyLaboratory RAMSES, Research & Innovation Technology Department, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy1st Orthopaedic and Traumatologic Clinic, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, ItalyUnit of Histology, Embryology and Applied Biology, Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via Belmeloro 8, 40126 Bologna, ItalyUnit of Histology, Embryology and Applied Biology, Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via Belmeloro 8, 40126 Bologna, ItalyUnit of Histology, Embryology and Applied Biology, Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via Belmeloro 8, 40126 Bologna, ItalyStem Sel Ltd., Viale Giuseppe Fanin 48, 40127 Bologna, ItalyStem Sel Ltd., Viale Giuseppe Fanin 48, 40127 Bologna, ItalyStem Sel Ltd., Viale Giuseppe Fanin 48, 40127 Bologna, ItalyMesenchymal stem cells (MSC) make up less than 1% of the bone marrow (BM). Several methods are used for their isolation such as gradient separation or centrifugation, but these methodologies are not direct and, thus, plastic adherence outgrowth or magnetic/fluorescent-activated sorting is required. To overcome this limitation, we investigated the use of a new separative technology to isolate MSCs from BM; it label-free separates cells based solely on their physical characteristics, preserving their native physical properties, and allows real-time visualization of cells. BM obtained from patients operated for osteochondral defects was directly concentrated in the operatory room and then analyzed using the new technology. Based on cell live-imaging and the sample profile, it was possible to highlight three fractions (F1, F2, F3), and the collected cells were evaluated in terms of their morphology, phenotype, CFU-F, and differentiation potential. Multipotent MSCs were found in F1: higher CFU-F activity and differentiation potential towards mesenchymal lineages compared to the other fractions. In addition, the technology depletes dead cells, removing unwanted red blood cells and non-progenitor stromal cells from the biological sample. This new technology provides an effective method to separate MSCs from fresh BM, maintaining their native characteristics and avoiding cell manipulation. This allows selective cell identification with a potential impact on regenerative medicine approaches in the orthopedic field and clinical applications.https://www.mdpi.com/2306-5354/9/2/49mesenchymal stem cellsbone marrowlabel-free separationcell selectionosteoarticular regeneration |
| spellingShingle | Silvia Zia Carola Cavallo Ilaria Vigliotta Valentina Parisi Brunella Grigolo Roberto Buda Pasquale Marrazzo Francesco Alviano Laura Bonsi Andrea Zattoni Pierluigi Reschiglian Barbara Roda Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples Bioengineering mesenchymal stem cells bone marrow label-free separation cell selection osteoarticular regeneration |
| title | Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples |
| title_full | Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples |
| title_fullStr | Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples |
| title_full_unstemmed | Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples |
| title_short | Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples |
| title_sort | effective label free sorting of multipotent mesenchymal stem cells from clinical bone marrow samples |
| topic | mesenchymal stem cells bone marrow label-free separation cell selection osteoarticular regeneration |
| url | https://www.mdpi.com/2306-5354/9/2/49 |
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