Long term tracking of mesenchymal stem cells differentiation to adipocytes with photostable fluorescent biodegradable nanoparticles
Date of Issue2015
School of Chemical and Biomedical Engineering
Many scientists explore the promising potential of stem cell for future medicine, leveraging from self-renewal and differentiation properties of stem cell. However, recent clinical studies showed conflicting results, due to lack of understanding in the distribution, homing process, engraftment, survival, and fate of stem cells after injection to application site in vivo. Fluorescent labeling of stem cell, being affordable and easily accessible, rises as one of prospective methods to overcome those difficulties. This study aims to examine the effect of cellular uptake of a novel fluorescent biodegradable polymeric nanoparticle on mesenchymal stem cells (MSC) for long-term stem cell tracking. After MSC was incubated with nanoparticles for 3 days and 7 days of uptake, cellular uptake was analyzed by confocal laser scanning microscopy (CLSM) and flow cytometry. Influence on cell viability and proliferation was analyzed by PrestoBlue® assay. Adipogenic differentiation potential was examined by quantifying marker genes by real-time RT-PCR and histology staining. Confocal microscopy and flow cytometry results demonstrated successful nanoparticle uptake and retention up to 21 days after differentiation induction. Cell viability and proliferation were unaffected. Adipogenic differentiation capability was found to be unaffected qualitatively and quantitatively. Thus, this novel nanoparticle has a strong foundation to be potential fluorescent label for long-term tracking of MSC during adipogenic differentiation. Future studies can extend to other differentiated lineages of MSC and other types of stem cell for a thorough performance analysis.
Final Year Project (FYP)
Nanyang Technological University