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GDNF Lentivirus Infection of Cynomolgus Monkey Bone Marrow Mesenchymal Stem Cells
Yu-Lan Wang1, Yan-Ling Xu2, Zhen-Hua Ren2,3*
1Central Laboratory, Xuanwu Hospital, Capital Medical University, Beijing 100053, China;2Cell Biology Laboratory, Xuanwu Hospital, Capital Medical University, Beijing 100053, China;3Department of Anatomy, Anhui Medical Uni
Abstract: Mesenchymal stem cells (MSCs) have received considerable attention for various therapeutic approaches in recent years. MSCs are also easy to genetically modify to express therapeutic genes using lentiviral vectors. In this study， we transfected cynomolgus monkey mesenchymal stem cells (MSCs) using lentiviral vectors encoding glial cell line derived neurotrophic factor (GDNF) to study its expression level in vitro. First， Lentiviral vectors encoding GDNF were packaged by 293T cells through three plasmids transient co-transfection method using standard lipofectamine reagent. The viral titers were tested by the transfection efficiency of 293T cells. At the same time， MSCs were transfected under different multiplicity of infection. GDNF gene expression level and protein secretion level of MSCs were tested by real-time PCR and ELISA methods after transfection. The results indicated lentiviral vectors encoding GDNF successfully transfected cynomolgus monkey MSCs， and over-expressed GDNF in vitro for over 1 month. Furthermore， GDNF expression was influenced by the multiplicity of infection. It might provide technical assistance for cynomolgus monkey MSCs and GDNF as gene therapy applied to the treatment of neurodegenerative diseases.
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