Estimating transfection efficiency in differentiated and undifferentiated neural cells

• Published in: BMC research notes Vol. 12; no. 1; pp. 225 - 7
• Main Authors: Alabdullah, Abeer A., Al-Abdulaziz, Basma, Alsalem, Hanan, Magrashi, Amna, Pulicat, Subramanian M., Almzroua, Amer A., Almohanna, Falah, Assiri, Abdullah Mohamed, Al Tassan, Nada A., Al-Mubarak, Bashayer R.
• Format: Journal Article
• Language: English
• Published: London BioMed Central 15.04.2019; BioMed Central Ltd; BMC
• Subjects: Animals; Antigens, Nuclear - genetics; Antigens, Nuclear - metabolism; Astrocytes; Astrocytes - cytology; Astrocytes - drug effects; Astrocytes - metabolism; Binding sites; Biomarkers - metabolism; Biomedical and Life Sciences; Biomedicine; Calcium phosphates; Cell Differentiation; Cell Line, Tumor; Cell lines; Cell Survival - drug effects; Cerebral Cortex - cytology; Cerebral Cortex - metabolism; Coculture Techniques; Cortex; Cytological research; Deoxyribonucleic acid; DNA; Efficiency; Gene Expression; Gene silencing; Genes, Reporter; Glial Fibrillary Acidic Protein - genetics; Glial Fibrillary Acidic Protein - metabolism; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Hippocampus; Humans; Life Sciences; Lipids - chemistry; Lipids - pharmacology; Lipofection; Medicine/Public Health; Methods; Mice; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neural Stem Cells - cytology; Neural Stem Cells - drug effects; Neural Stem Cells - metabolism; Neuroblastoma; Neuroblastoma cell lines; Neurons; Neurons - cytology; Neurons - drug effects; Neurons - metabolism; Organ Specificity; Physiological aspects; Plasmids - chemistry; Plasmids - metabolism; Primary Cell Culture; Primary cortical astrocytes; Primary cortical neurons; Research Note; Transfection; Transfection - methods; Transfection efficiency; Vectors (Biology); Saudi Arabia; Primary cortical neurons; Lipofection; Primary cortical astrocytes; Neuroblastoma cell lines; Transfection efficiency
• ISSN: 1756-0500; 1756-0500
• DOI: 10.1186/s13104-019-4249-5

Objective Delivery of constructs for silencing or over-expressing genes or their modified versions is a crucial step for studying neuronal cell biology. Therefore, efficient transfection is important for the success of these experimental techniques especially in post-mitotic cells like neurons. In this study, we have assessed the transfection rate, using a previously established protocol, in both primary cortical cultures and neuroblastoma cell lines. Transfection efficiencies in these preparations have not been systematically determined before. Results Transfection efficiencies obtained herein were (10–12%) for neuroblastoma, (5–12%) for primary astrocytes and (1.3–6%) for primary neurons. We also report on cell-type specific transfection efficiency of neurons and astrocytes within primary cortical cultures when applying cell-type selective transfection protocols. Previous estimations described in primary cortical or hippocampal cultures were either based on general observations or on data derived from unspecified number of biological and/or technical replicates. Also to the best of our knowledge, transfection efficiency of pure primary neuronal cultures or astrocytes cultured in the context of pure or mixed (neurons/astrocytes) population cultures have not been previously determined. The transfection strategy used herein represents a convenient, and a straightforward tool for targeted cell transfection that can be utilized in a variety of in vitro applications.