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Estimating transfection efficiency in differentiated and undifferentiated neural cells

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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466792/
https://www.ncbi.nlm.nih.gov/pubmed/30987672
http://dx.doi.org/10.1186/s13104-019-4249-5
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author 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.
author_facet 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.
author_sort Alabdullah, Abeer A.
collection PubMed
description 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-019-4249-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-64667922019-04-22 Estimating transfection efficiency in differentiated and undifferentiated neural cells 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. BMC Res Notes Research Note 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-019-4249-5) contains supplementary material, which is available to authorized users. BioMed Central 2019-04-15 /pmc/articles/PMC6466792/ /pubmed/30987672 http://dx.doi.org/10.1186/s13104-019-4249-5 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Note
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.
Estimating transfection efficiency in differentiated and undifferentiated neural cells
title Estimating transfection efficiency in differentiated and undifferentiated neural cells
title_full Estimating transfection efficiency in differentiated and undifferentiated neural cells
title_fullStr Estimating transfection efficiency in differentiated and undifferentiated neural cells
title_full_unstemmed Estimating transfection efficiency in differentiated and undifferentiated neural cells
title_short Estimating transfection efficiency in differentiated and undifferentiated neural cells
title_sort estimating transfection efficiency in differentiated and undifferentiated neural cells
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466792/
https://www.ncbi.nlm.nih.gov/pubmed/30987672
http://dx.doi.org/10.1186/s13104-019-4249-5
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