Estimating transfection efficiency in differentiated and undifferentiated neural cells

Abstract

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.

SUMOylation regulates protein cargo in Astrocyte-derived small extracellular vesicles

“…This may be due to the low levels of transfection that we obtained in astrocytes, of about 15%. This is consistent with publications that describe the low transfection efficiency of astrocytes (Alabdullah et al, 2019), but also, that SUMOylation is a transient event that does not increase steady state levels (Henley, Craig, & Wilkinson, 2014;Klug et al, 2013).…”

Non-canonical Keap1-independent activation of Nrf2 in astrocytes by mild oxidative stress

“…Astrocytes within mixed astrocyte/neuronal cultures were transfected with Neh2-luc ( B ) or control luciferase reporter plasmid ( C ) plus a pTK-renilla plasmid. The approach to target astrocyte transfection within mixed neuronal/astrocyte cultures was described previously [ 38 , 42 , 43 ] and verified for this study ( Figs. S3a and b ).…”

Presenilin 1 Regulates [Ca2+]i and Mitochondria/ER Interaction in Cultured Rat Hippocampal Neurons

“…In both groups of neurons, there was a nearly complete overlap of the transfected PS1/mPS1 and the immunostaining for PS1 (Supplementary Figure 1) indicating that the antibody recognizes both transfected varieties of PS1. However, despite extensive experience with transfection methodologies, there is still a paucity of information on either the selective vulnerability of a small proportion of the neurons to transfection as well as the cellular impact of the transfection (see [21]).…”