![]() Some studies have suggested that cell transfection with fluorescent proteins 5, 6, 7 will be required for long term cell tracking and others are pursuing magnetic nanoparticles 8, bioluminescent probes 9, 10, 11, quantum dots 12, 13 and radioactive isotope alternatives 14 for stem cell labeling. To this end, a number of cell labeling techniques have been developed. A tool to longitudinally follow MSC in 3D is essential to better understand cell migration and organization in 3D microenvironments. Regardless of the approach, to improve the effectiveness of such applications it is critical to understand the determinants of stem cell migration in 3D microenvironments.ĭespite the worldwide effort to modulate and direct stem cell fate for tissue regeneration, the influence of 3D extracellular matrix (ECM) features on MSC motility still remains largely unknown. Most strategies being currently explored involve either stem cell transplantation to patients 1, implantation of the triad scaffolds/stem cells/growth factors 2, 3, or the use of materials that stimulate endogenous stem cell recruitment 4. MSC are an attractive cell source for regenerative cell-based therapies due to its well established multipotency, immunomodulatory and paracrine properties, combined with their ability to migrate into damaged tissues. This platform thus provides a straightforward approach to characterize MSC dynamics in 3D and has applications in the field of stem cell biology and for the development of biomaterials for tissue regeneration. Comparison of cells mobility within matrices with tuned physicochemical properties revealed that MSC embedded in Matrigel migrated 64% more with 5.2 mg protein/mL than with 9.6 mg/mL and that MSC embedded in RGD-alginate migrated 51% faster with 1% polymer concentration than in 2% RGD-alginate. This strategy provided reliable tracking in 3D microenvironments with different properties, including the hydrogels Matrigel and alginate as well as chitosan porous scaffolds. Human MSC were transfected to express a fluorescent photoswitchable protein, Dendra2, which was used to highlight and follow the same group of cells for more than seven days, even if removed from the microscope to the incubator. Here, a simple and reliable imaging technique was developed to study MSC dynamical behavior in natural and bioengineered 3D matrices. However, the identity and plasticity of skeletal stem and progenitor cells in vivo is still unclear and could be further defined using lineage tracing with additional markers or marker combinations.Mesenchymal Stem/Stromal Cells (MSC) are a promising cell type for cell-based therapies - from tissue regeneration to treatment of autoimmune diseases - due to their capacity to migrate to damaged tissues, to differentiate in different lineages and to their immunomodulatory and paracrine properties. Lineage tracing has been used widely used to identify chondro- and osteoprogenitors during embryonic development, postnatal growth, fracture or bone injury, and heterotopic ossification. Lineage tracing is performed as a “pulse–chase” experiment, with characterization of the initially labeled population being critical for interpretation of the resulting chase. This allows indefinite labeling of cells with a visual marker following induction of Cre activity. Genetic lineage tracing is generally performed by combining an inducible Cre expressed in a cell type of interest with a Cre-activated reporter usually expressing a fluorescent protein. Lineage tracing is a technique used to identify and track cell populations in vivo. ![]()
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