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Volume 9
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10.3390/bioengineering9020049
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Article

Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples

by
Silvia Zia
1,†,
Carola Cavallo
2,†,
Ilaria Vigliotta
3,
Valentina Parisi
2,
Brunella Grigolo
2,
Roberto Buda
4,
Pasquale Marrazzo
3,
Francesco Alviano
3,
Laura Bonsi
3,
Andrea Zattoni
1,5,
Pierluigi Reschiglian
1,5 and
Barbara Roda
1,5,*
1
Stem Sel Ltd., Viale Giuseppe Fanin 48, 40127 Bologna, Italy
2
Laboratory RAMSES, Research & Innovation Technology Department, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy
3
Unit of Histology, Embryology and Applied Biology, Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via Belmeloro 8, 40126 Bologna, Italy
4
1st Orthopaedic and Traumatologic Clinic, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy
5
Department of Chemistry “G. Ciamician”, University of Bologna, Via Selmi 2, 40126 Bologna, Italy
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Bioengineering 2022, 9(2), 49; https://doi.org/10.3390/bioengineering9020049
Submission received: 29 December 2021 / Revised: 14 January 2022 / Accepted: 17 January 2022 / Published: 22 January 2022
(This article belongs to the Special Issue Stem Cell Bioprocessing and Manufacturing, Volume 2)
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Abstract

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.
Keywords: mesenchymal stem cells; bone marrow; label-free separation; cell selection; osteoarticular regeneration mesenchymal stem cells; bone marrow; label-free separation; cell selection; osteoarticular regeneration

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MDPI and ACS Style

Zia, S.; Cavallo, C.; Vigliotta, I.; Parisi, V.; Grigolo, B.; Buda, R.; Marrazzo, P.; Alviano, F.; Bonsi, L.; Zattoni, A.; et al. Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples. Bioengineering 2022, 9, 49. https://doi.org/10.3390/bioengineering9020049

AMA Style

Zia S, Cavallo C, Vigliotta I, Parisi V, Grigolo B, Buda R, Marrazzo P, Alviano F, Bonsi L, Zattoni A, et al. Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples. Bioengineering. 2022; 9(2):49. https://doi.org/10.3390/bioengineering9020049

Chicago/Turabian Style

Zia, Silvia, Carola Cavallo, Ilaria Vigliotta, Valentina Parisi, Brunella Grigolo, Roberto Buda, Pasquale Marrazzo, Francesco Alviano, Laura Bonsi, Andrea Zattoni, and et al. 2022. "Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples" Bioengineering 9, no. 2: 49. https://doi.org/10.3390/bioengineering9020049

APA Style

Zia, S., Cavallo, C., Vigliotta, I., Parisi, V., Grigolo, B., Buda, R., Marrazzo, P., Alviano, F., Bonsi, L., Zattoni, A., Reschiglian, P., & Roda, B. (2022). Effective Label-Free Sorting of Multipotent Mesenchymal Stem Cells from Clinical Bone Marrow Samples. Bioengineering, 9(2), 49. https://doi.org/10.3390/bioengineering9020049

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