Differentiation of genetically modified canine bone mesenchymal stem cells labeled with superparamagnetic iron oxide
Xing-Long Liu1, Qing-Quan Zu1, Bin Wang1
1Radiology Department, The First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, Jiangsu 210029, P.R. China.
Abstract:
The use of mesenchymal stem cells (MSCs) has been reported to improve outcomes in various types of nervous system diseases, primarily based on their neural regenerative differentiation ability and paracrine effect on different neuroprotective cytokines. Genetically modified MSCs may enhance the paracrine effect and may further improve the cell‑based therapeutic outcome of nervous system diseases. Magnetic resonance imaging has been used to monitor distribution and migration of cells labeled with superparamagnetic iron oxide (SPIO) nanoparticles. However, few studies have described the neural differentiation ability of genetically modified and SPIO‑labeled MSCs, which is the foundation for cell tracking and cell therapy in vivo. In this study, canine bone marrow‑derived MSCs (BMSCs) were initially labeled with SPIO, by culturing with 20 µg/ml SPIO for 24 h, and transfected with the brain‑derived neurotrophic factor (BDNF) gene using lentivirus transfection at different multiplicities of infection (MOI) values. The optimized MOI value was demonstrated by cellular viability and enhanced green fluorescent protein (eGFP) rate. Subsequently, the BMSCs were induced to differentiate into neuron‑like cells by chemical induction. The results demonstrated that BDNF‑overexpressing BMSCs labeled with SPIO can be induced into neuron‑like cells with high efficiency and minimal effects on cell viability. Additionally, following neural differentiation, the cells transfected with BDNF and labeled with SPIO expressed significantly higher levels of BDNF and neural markers. The overexpression of BDNF may contribute to neural differentiation of BDNFs, and may have potential benefits for further BMSC‑based therapy in vivo.
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