Blog
About

0
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Sorting living mesenchymal stem cells using a TWIST1 RNA-based probe depends on incubation time and uptake capacity

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Bone marrow derived mesenchymal stromal cells (BMSCs) are multipotent progenitors of particular interest for cell-based tissue engineering therapies. However, one disadvantage that limit their clinical use is their heterogeneity. In the last decades a great effort was made to select BMSC subpopulations based on cell surface markers, however there is still no general consensus on which markers to use to obtain the best BMSCs for tissue regeneration. Looking for alternatives we decided to focus on a probe-based method to detect intracellular mRNA in living cells, the SmartFlare technology. This technology does not require fixation of the cells and allows us to sort living cells based on gene expression into functionally different populations. However, since the technology is available it is debated whether the probes specifically recognize their target mRNAs. We validated the TWIST1 probe and demonstrated that it specifically recognizes TWIST1 in BMSCs. However, differences in probe concentration, incubation time and cellular uptake can strongly influence signal specificity. In addition we found that TWIST1 high expressing cells have an increased expansion rate compared to TWIST1 low expressing cells derived from the same initial population of BMSCs. The SmartFlare probes recognize their target gene, however for each probe and cell type validation of the protocol is necessary.

          Electronic supplementary material

          The online version of this article (10.1007/s10616-019-00355-w) contains supplementary material, which is available to authorized users.

          Related collections

          Most cited references 30

          • Record: found
          • Abstract: found
          • Article: not found

          Multilineage potential of adult human mesenchymal stem cells.

          Human mesenchymal stem cells are thought to be multipotent cells, which are present in adult marrow, that can replicate as undifferentiated cells and that have the potential to differentiate to lineages of mesenchymal tissues, including bone, cartilage, fat, tendon, muscle, and marrow stroma. Cells that have the characteristics of human mesenchymal stem cells were isolated from marrow aspirates of volunteer donors. These cells displayed a stable phenotype and remained as a monolayer in vitro. These adult stem cells could be induced to differentiate exclusively into the adipocytic, chondrocytic, or osteocytic lineages. Individual stem cells were identified that, when expanded to colonies, retained their multilineage potential.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment.

            The identity of cells that establish the hematopoietic microenvironment (HME) in human bone marrow (BM), and of clonogenic skeletal progenitors found in BM stroma, has long remained elusive. We show that MCAM/CD146-expressing, subendothelial cells in human BM stroma are capable of transferring, upon transplantation, the HME to heterotopic sites, coincident with the establishment of identical subendothelial cells within a miniature bone organ. Establishment of subendothelial stromal cells in developing heterotopic BM in vivo occurs via specific, dynamic interactions with developing sinusoids. Subendothelial stromal cells residing on the sinusoidal wall are major producers of Angiopoietin-1 (a pivotal molecule of the HSC "niche" involved in vascular remodeling). Our data reveal the functional relationships between establishment of the HME in vivo, establishment of skeletal progenitors in BM sinusoids, and angiogenesis.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: found
              Is Open Access

              Aging of mesenchymal stem cell in vitro

              Background A hot new topic in medical treatment is the use of mesenchymal stem cells (MSC) in therapy. The low frequency of this subpopulation of stem cells in bone marrow (BM) necessitates their in vitro expansion prior to clinical use. We evaluated the effect of long term culture on the senescence of these cells. Results The mean long term culture was 118 days and the mean passage number was 9. The average number of PD decreased from 7.7 to 1.2 in the 10th passage. The mean telomere length decreased from 9.19 Kbp to 8.7 kbp in the 9th passage. Differentiation potential dropped from the 6th passage on. The culture's morphological abnormalities were typical of the Hayflick model of cellular aging. Conclusion We believe that MSC enter senescence almost undetectably from the moment of in vitro culturing. Simultaneously these cells are losing their stem cell characteristics. Therefore, it is much better to consider them for cell and gene therapy early on.
                Bookmark

                Author and article information

                Contributors
                r.narcisi@erasmusmc.nl
                Journal
                Cytotechnology
                Cytotechnology
                Cytotechnology
                Springer Netherlands (Dordrecht )
                0920-9069
                1573-0778
                14 November 2019
                14 November 2019
                February 2020
                : 72
                : 1
                : 37-45
                Affiliations
                [1 ]GRID grid.5645.2, ISNI 000000040459992X, Department of Orthopaedics, , Erasmus MC, ; 3015 CN Rotterdam, The Netherlands
                [2 ]GRID grid.5645.2, ISNI 000000040459992X, Department of Internal Medicine, , Erasmus MC, ; Rotterdam, The Netherlands
                [3 ]GRID grid.5606.5, ISNI 0000 0001 2151 3065, Department of Experimental Medicine, , University of Genova, ; Genoa, Italy
                [4 ]GRID grid.5645.2, ISNI 000000040459992X, Department of Otorhinolaryngology, , Erasmus MC, ; 3015 CN Rotterdam, The Netherlands
                Article
                355
                10.1007/s10616-019-00355-w
                7002702
                31728801
                © The Author(s) 2019

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.

                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100006315, ReumaNederland;
                Award ID: 16-1-201
                Award Recipient :
                Funded by: Veni by NWO
                Award ID: 13659
                Award Recipient :
                Categories
                Original Article
                Custom metadata
                © Springer Nature B.V. 2020

                Comments

                Comment on this article