Author/Authors :
Mortazavi، Yousef نويسنده , , Sheikhsaran، Fatemeh نويسنده Department of Molecular Medicine and Genetics, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran , , Khamisipour، Gholamreza نويسنده Hematology Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran , , Soleimani، Masoud نويسنده , , Teimuri، Ali نويسنده Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran , , Shokri، Somayeh نويسنده Department of Molecular Medicine and Genetics, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran ,
Abstract :
Objective: Treatment and repair of neurodegenerative diseases such as brain tumors,
spinal cord injuries, and functional disorders, including Alzheimer’s disease, are challenging
problems. A common treatment approach for such disorders involves the use of
mesenchymal stem cells (MSCs) as an alternative cell source to replace injured cells.
However, use of these cells in hosts may potentially cause adverse outcomes such as tumorigenesis
and uncontrolled differentiation. In attempt to generate mesenchymal derived
neural cells, we have infected MSCs with recombinant lentiviruses that expressed nerve
growth factor (NGF) and assessed their neural lineage genes.
Materials and Methods: In this experimental study, we cloned the NGF gene sequence
into a helper dependent lentiviral vector that contained the green fluorescent protein (GFP)
gene. The recombinant vector was amplified in DH5 bacterial cells. Recombinant viruses
were generated in the human embryonic kidney 293 (HEK-293) packaging cell line with
the helper vectors and analyzed under fluorescent microscopy. Bone marrow mesenchymal
cells were infected by recombinant viruses for three days followed by assessment of
neural differentiation. We evaluated expression of NGF through measurement of the NGF
protein in culture medium by ELISA; neural specific genes were quantified by real-time
polymerase chain reaction (PCR).
Results: We observed neural morphological changes after three days. Quantitative PCR
showed that expressions of NESTIN, glial derived neurotrophic factor (GDNF), glial fibrillary
acidic protein (GFAP) and Microtubule-associated protein 2 (MAP2) genes increased
following induction of NGF overexpression, whereas expressions of endogenous NGF
and brain derived neural growth factor (BDNF) genes reduced.
Conclusion: Ectopic expression of NGF can induce neurogenesis in MSCs. Direct injection
of MSCs may cause tumorigenesis and an undesirable outcome. Therefore an
alternative choice to overcome this obstacle may be the utilization of differentiated neural
stem cells.
