Estimating transfection efficiency in differentiated and undifferentiated neural cells

Autor: Bashayer R. Al-Mubarak, Amer A. Almzroua, Nada A. Al Tassan, Abdullah Mohamed Assiri, Falah Al-Mohanna, Abeer A. Alabdullah, Hanan Alsalem, Basma S. AlAbdulaziz, Amna Magrashi, Subramanian M. Pulicat
Rok vydání: 2019
Předmět:
0301 basic medicine
viruses
Gene Expression
lcsh:Medicine
Hippocampal formation
Mice
0302 clinical medicine
Neural Stem Cells
Genes
Reporter
030212 general & internal medicine
lcsh:QH301-705.5
Lipofection
Cerebral Cortex
Neurons
education.field_of_study
Chemistry
Antigens
Nuclear
Cell Differentiation
Transfection efficiency
General Medicine
Transfection
Lipids
Cell biology
Research Note
Organ Specificity
embryonic structures
Microtubule-Associated Proteins
Plasmids
animal structures
Cell Survival
Green Fluorescent Proteins
Primary Cell Culture
Population
Nerve Tissue Proteins
Context (language use)
Neuroblastoma cell lines
Primary cortical neurons
General Biochemistry
Genetics and Molecular Biology
03 medical and health sciences
Cell Line
Tumor
Neuroblastoma
Glial Fibrillary Acidic Protein
medicine
Animals
Humans
Gene silencing
lcsh:Science (General)
education
Gene
lcsh:R
fungi
medicine.disease
Primary cortical astrocytes
Coculture Techniques
In vitro
030104 developmental biology
lcsh:Biology (General)
Astrocytes
Biomarkers
lcsh:Q1-390
Zdroj: BMC Research Notes, Vol 12, Iss 1, Pp 1-7 (2019)
BMC Research Notes
ISSN: 1756-0500
DOI: 10.1186/s13104-019-4249-5
Popis: 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.
Databáze: OpenAIRE
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