Estimating transfection efficiency in differentiated and undifferentiated neural cells.

Basma Al-Abdulaziz
Basma Al-Abdulaziz
King Faisal Specialist Hospital and Research Center
Hanan Alsalem
Hanan Alsalem
King Faisal Specialist Hospital and Research Center
Amna Magrashi
Amna Magrashi
King Faisal Specialist Hospital & Research Center
Falah Almohanna
Falah Almohanna
King Faisal Specialist Hospital and Research Centre

BMC Res Notes 2019 Apr 15;12(1):225. Epub 2019 Apr 15.

Behavioral Genetics Unit, Department of Genetics, King Faisal Specialist Hospital and Research Center, Riyadh, 11211, Saudi Arabia.

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.

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April 2019
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