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Physiology and Pharmacology، جلد ۲۷، شماره ۱، صفحات ۰-۰
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چکیده فارسی مقاله
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عنوان انگلیسی Efficient Modified-mRNA Transfection in Neural Stem Cells
چکیده انگلیسی مقاله Background: Neural stem cells (NSCs) are multipotent stem cells residing in the central nervous system that are capable of self-renewal to support ongoing requirements for neurogenesis in the adult brain. Since they are considered potential candidate cells for neuro-regenerative medicine, applying safe induction methods for them is very important. Synthetic modified-mRNA (mmRNA) as an alternative to traditional DNA- or protein-based methods, is regarded as a powerful tool for inducing short-term gene expression in cells with no genetic manipulation. Methods: Here, we aimed to develop an optimized condition for mmRNA transfection in primary NSCs. In vitro-transcribed EGFP mmRNA (mmRNAEGFP) was delivered to human embryonic kidney cells (HEK293T) and mouse NSCs by using two commercial agents, Lipofectamine-2000 (LF2000) and TransIT. Also, a plasmid DNA was used to transfect cells considered EGFP-expressing positive control. In addition, the poly(A) tail (poly adenosine tail) elongation and chloroquine (CQ) treatment were performed to improve transfection efficiency. Finally, flow cytometry, fluorescence microscopy and MTT assays were performed to assess the cells. Results: In comparison with HEK293T, NSCs were very sensitive to transfection, the efficacy of transfection using DNA/LF2000 was higher in HEK293T cells, but mmRNAEGFP/TransIT showed better transfection efficacy in NSCs. Poly(A) tail elongation; also, treating the cells with CQ prior to transfection significantly improved its efficacy. Conclusions: The mmRNA poly(A) tail elongation, and the use of specific transfection agents in combination with TLR inhibitors can lead to a more effective transfection in NSCs.
کلیدواژه‌های انگلیسی مقاله Modified-mRNA, In vitro transcription, Neural stem cell, Chloroquine, Transfection, Cell culture
نویسندگان مقاله | Reyhaneh Khayamabed
Department of Biology, ACECR Institute of Higher Education, Isfahan, Iran


| Naeimeh Rezaie
Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran


| Elnaz Poorgolizadeh
Department of Biology, ACECR Institute of Higher Education, Isfahan, Iran


| Farshad Homayouni Moghadam
Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran


| Kianoush Dorminani
Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran


| Mohammad Hossein Nasr Esfahani
Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran
نشانی اینترنتی http://ppj.phypha.ir/browse.php?a_code=A-10-146-2&slc_lang=en&sid=1
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کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده Cell, Stem Cell and Cancer
نوع مقاله منتشر شده Experimental research article
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