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      Differentiation of single lymphoma primary cells and normal B-cells based on their adhesion to mesenchymal stromal cells in optical tweezers

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          Abstract

          We have adapted a non-invasive method based on optical tweezers technology to differentiate between the normal B-cells and the B-cell non-Hodgkin lymphoma (B-NHL) cells derived from clinical samples. Our approach bases on the nascent adhesion between an individual B-cell and a mesenchymal stromal cell. In this study, a single B-cell was trapped and optically seeded on a mesenchymal stromal cell and kept in a direct contact with it until a stable connection between the cells was formed in time scale. This approach allowed us to avoid the introduction of any exogenous beads or chemicals into the experimental setup which would have affected the cell-to-cell adhesion. Here, we have provided new evidence that aberrant adhesive properties found in transformed B-cells are related to malignant neoplasia. We have demonstrated that the mean time required for establishing adhesive interactions between an individual normal B-cell and a mesenchymal stromal cell was 26.7 ± 16.6 s, while for lymphoma cell it was 208.8 ± 102.3 s, p < 0.001. The contact time for adhesion to occur ranged from 5 to 90 s and from 60 to 480 s for normal B-cells and lymphoma cells, respectively. This method for optically controlled cell-to-cell adhesion in time scale is beneficial to the successful differentiation of pathological cells from normal B-cells within the fine needle aspiration biopsy of a clinical sample. Additionally, variations in time-dependent adhesion among subtypes of B-NHL, established here by the optical trapping, confirm earlier results pertaining to cell heterogeneity.

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          Observation of a single-beam gradient force optical trap for dielectric particles

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            Immunosuppressive effect of mesenchymal stem cells favors tumor growth in allogeneic animals.

            Mesenchymal stem cells (MSCs) are largely studied for their potential clinical use. Recently, they have gained further interest after demonstration of an immunosuppressive role. In this study, we investigated whether in vivo injection of MSCs could display side effects related to systemic immunosuppression favoring tumor growth. We first showed in vitro that the murine C3H10T1/2 (C3) MSC line and primary MSCs exhibit immunosuppressive properties in mixed lymphocyte reaction. We demonstrated that this effect is mediated by soluble factors, secreted only on "activation" of MSCs in the presence of splenocytes. Moreover, the immunosuppression is mediated by CD8+ regulatory cells responsible for the inhibition of allogeneic lymphocyte proliferation. We then demonstrated that the C3 MSCs expressing the human bone morphogenetic protein 2 (hBMP-2) differentiation factor were not rejected when implanted in various allogeneic immunocompetent mice and were still able to differentiate into bone. Importantly, using a murine melanoma tumor model, we showed that the subcutaneous injection of B16 melanoma cells led to tumor growth in allogeneic recipients only when MSCs were coinjected. Although the potential side effects of immunosuppression induced by MSCs have to be considered in further clinical studies, the usefulness of MSCs for various therapeutic applications still remains of great interest.
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              Optical tweezers for single cells.

              Optical tweezers (OT) have emerged as an essential tool for manipulating single biological cells and performing sophisticated biophysical/biomechanical characterizations. Distinct advantages of using tweezers for these characterizations include non-contact force for cell manipulation, force resolution as accurate as 100aN and amiability to liquid medium environments. Their wide range of applications, such as transporting foreign materials into single cells, delivering cells to specific locations and sorting cells in microfluidic systems, are reviewed in this article. Recent developments of OT for nanomechanical characterization of various biological cells are discussed in terms of both their theoretical and experimental advancements. The future trends of employing OT in single cells, especially in stem cell delivery, tissue engineering and regenerative medicine, are prospected. More importantly, current limitations and future challenges of OT for these new paradigms are also highlighted in this review.
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                Author and article information

                Contributors
                kamila.dus-szachniewicz@umed.wroc.pl
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                8 July 2019
                8 July 2019
                2019
                : 9
                : 9885
                Affiliations
                [1 ]ISNI 0000 0001 1090 049X, GRID grid.4495.c, Department of Pathology, , Wrocław Medical University, ; Marcinkowskiego 1, 50-368 Wrocław, Poland
                [2 ]ISNI 0000 0000 9805 3178, GRID grid.7005.2, Department of Optics and Photonics, , Wrocław University of Science and Technology, Faculty of Fundamental Problems of Technology, ; Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland
                [3 ]ISNI 0000 0001 0347 9385, GRID grid.13252.37, Department of Statistics, , Wrocław University of Economics, ; Komandorska 118/120, 53-345 Wrocław, Poland
                [4 ]ISNI 0000 0001 1090 049X, GRID grid.4495.c, 2nd Department of General and Oncological Surgery, , Wrocław Medical University, ; Borowska 213, 50-556 Wrocław, Poland
                [5 ]Division of Pathology, Sokołowski Hospital Wałbrzych, Sokołowskiego 4, 58-309 Wałbrzych, Poland
                [6 ]Flow Cytometry Laboratory, Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Institute-Oncology Centre, Wilhelma Konrada Roentgena 5, 02-781 Warsaw, Poland
                Author information
                http://orcid.org/0000-0001-9005-9695
                Article
                46086
                10.1038/s41598-019-46086-y
                6614388
                31285461
                b0db570e-701d-4262-87da-ca1588a7c633
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 5 March 2019
                : 21 June 2019
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100005632, Ministry of Science and Higher Education | Narodowe Centrum Bada&amp;#x0144; i Rozwoju (National Centre for Research and Development);
                Award ID: LIDER/016/275/L-5/13/NCBR/2014
                Award ID: LIDER/016/275/L-5/13/NCBR/2014
                Award ID: LIDER/016/275/L-5/13/NCBR/2014
                Award ID: LIDER/016/275/L-5/13/NCBR/2014
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100004281, Narodowe Centrum Nauki (National Science Centre);
                Award ID: UMO-2017/27/B/ST7/01255
                Award Recipient :
                Categories
                Article
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                © The Author(s) 2019

                Uncategorized
                b-cell lymphoma,optical manipulation and tweezers
                Uncategorized
                b-cell lymphoma, optical manipulation and tweezers

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