Neural Cell Transfection
Mature cultures of neurons and other neural cells are extremely valuable for in vitro neurotoxicity studies and screening for agents that can slow, stop, or even reverse the course of neurodegenerative diseases. The transfection of nucleic acids into neurons is essential for studying many aspects of neurobiology. However, neurons are among the most difficult cell types to achieve high transfection efficiency. They are very sensitive to culture conditions, presenting a particular challenge with regards to efficiencies. In addition to yielding low efficiencies, currently available cationic lipid reagents are often toxic to the cells, compromising post-transfection experimental results. While some viral mediated gene delivery systems have been shown to produce high efficiencies, they are very labor intensive and inconvenient for most researchers, along with the inherent danger and risk of provoking an immune response in the cell and/or interfering with the host genome. Using MTI-GlobalStem's neural cell transfection reagents, get the results needed with cost-effective reagents and easy to use protocols.
Figure 1. High GFP Expression in Primary Neurons– Primary rat cortical neurons were transfected with DNA-In® Neuro Transfection Reagent and incubated overnight in complete culture media. The above images were taken 48-hours post-transfection and show uniform, high GFP expression in healthy cells.
Figure 2. Superior mRNA Transfection efficiency in Primary Neurons – Primary rat cortical neurons cultured in 24-well plates were transfected with mRNA-In® Neuro Transfection Reagent and incubated overnight in complete culture media. Maximum transfection efficiency was maintained with reducing amounts of mRNA, from 500ng (high) to 125ng (low) amounts of mRNA. The above images were taken 48-hours post-transfection. In contrast to DNA transfection, neurons transfected with mRNA-In® Neuro showed uniform GFP expression from high (500ng) to very low (125ng) amounts of mRNA.
Figure 3. Neural progenitor cells (HT223-B) from adult brain were transfected with pEF1a-Cas9-2A-GFP plasmid (Addgene) using DNA-In® Stem (left) and Lipofectamine 3000 (right) . Data courtesy of Sasha Beylina, Staff Scientist, NIH/NIA
Figure. 4 Gene Editing by co-Transfection using EditPro™ Stem Transfection Reagent in Neural Stem Cells: iPSC- derived human neural stem cells were transfected in suspension using EditPro™ Stem with 250 ng Cas9 mRNA (modified)/ gRNA: Emx-1 crRNA (Exon2)-tracrRNA oligo/ GFP mRNA (modified) or Cas9 protein/gRNA: Emx-1 crRNA (Exon2)-tracrRNA oligo/ GFP mRNA (modified). GFP was observed after 24 hours. Genome-modification was analyzed using the T7Endo 1 assay.
|DNA-In® Neuro||Recommended for high efficiency DNA-delivery in difficult-to-transfect neurons with low cytotoxicity. Highly effective in cryopreserved and freshly isolated primary neuronal cells in vitro, as well as in iPSC-derived neurons.|
|DNA-In® Stem||Recommended for neural stem cell transfection applications requiring high efficiency DNA delivery.|
|DNA-In® SY5Y||Recommended for applications requiring high transfection efficiency in SH-SY5Y neuroblastoma cells. Optimized for high DNA delivery with exceptionally low cytotoxicity.|
|Ideal for neurobiology applications that require exceptionally high transfection efficiency and without the risk of genomic integration. mRNA-In Neuro is designed to provide exceptionally high transfection efficiency and use very low amounts of mRNA for maximum cell viability.|
|EditPro™ Stem||>90% transfection of NSC with DNA, RNA and/or protein. Ideal for geneome editing.|
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