In vitro non-viral murine pro-neurotrophin 3 gene transfer into rat bone marrow stromal cells

Abstract Neurotrophin 3 (NT-3) is an important factor for promoting prenatal neural development, as well as regeneration, axogenesis and plasticity in postnatal life. Therapy with NT-3 was reported to improve the condition of patients suffering from degenerative diseases and traumatic injuries, howe...

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Bibliographic Details
Published in:Journal of the neurological sciences 2017-04, Vol.375, p.137-145
Main Authors: Darabi, Shahram, Tiraihi, Taki, Delshad, AliReza, Sadeghizadeh, Majid, Khalil, Wisam, Taheri, Taher
Format: Article
Language:English
Subjects:
Animals
Antigens, CD - metabolism
BMSCs
Bone Marrow Cells - metabolism
Cells, Cultured
Genetic Vectors
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Mice
Neurology
Neurotrophin 3 - genetics
Neurotrophin 3 - metabolism
Non-viral transfection
NT-3
Rats
RNA, Messenger - metabolism
Secretory vector
Transfection
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Summary:Abstract Neurotrophin 3 (NT-3) is an important factor for promoting prenatal neural development, as well as regeneration, axogenesis and plasticity in postnatal life. Therapy with NT-3 was reported to improve the condition of patients suffering from degenerative diseases and traumatic injuries, however, the disadvantage of NT-3 protein delivery is its short half-life, thus our alternative approach is the use of NT-3 gene therapy. In this study, the bone marrow stromal cells (BMSCs) were isolated from adult rats, cultured for 4 passages and transfected with either pEGFP-N1 or a constructed vector containing murine proNT-3 (pSecTag2/HygroB-murine proNT-3) using Lipofectamine 2000 followed by Hygromycin B (200 mg/kg). The transfection efficiency of the transiently transfected BMSCs was evaluated using the green fluorescence protein containing vector (pEGFP-N1). A quantitative evaluation of the NT-3 expression of mRNA using real time QRT-PCR shows that there was double fold increase in NT-3 gene expression compared with non-transfected BMSCs, also, the culture supernatant yielded double fold increase in NT-3 using ELISA technique, the data were supported by immunoblotting technique. This suggests that the use of this transfection technique can be useful for gene therapy in different neurological disorders with neurodegenerative or traumatic origins.
ISSN:0022-510X
1878-5883
DOI:10.1016/j.jns.2017.01.058