Novel cell separation method for molecular analysis of neuron-astrocyte co-cultures
| العنوان: | Novel cell separation method for molecular analysis of neuron-astrocyte co-cultures |
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| المؤلفون: | Stéphane H. R. Oliet, Andrea Goudriaan, Mark H. G. Verheijen, Nutabi Camargo, Karen E. Carney, August B. Smit |
| المساهمون: | Molecular and Cellular Neurobiology, AIMMS, Neuroscience Campus Amsterdam - Brain Mechanisms in Health & Disease |
| المصدر: | Frontiers in Cellular Neuroscience Frontiers in Cellular Neuroscience, 8:12. Frontiers Media S.A. Frontiers in Cellular Neuroscience, Vol 8 (2014) Goudriaan, A, Camargo, N K, Carney, K E, Oliet, S H R, Smit, A B & Verheijen, M H G 2014, ' Novel cell separation method for molecular analysis of neuron-astrocyte co-cultures ', Frontiers in Cellular Neuroscience, vol. 8, 12, pp. 12 . https://doi.org/10.3389/fncel.2014.00012 |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | Messenger RNA, isolation approaches, astrocytes, RNA, Biology, co-culture, In vitro, lcsh:RC321-571, Cellular and Molecular Neuroscience, medicine.anatomical_structure, nervous system, Synaptic plasticity, medicine, Methods Article, method, transcriptional activation, Centrifugation, Neuron, neuron-glia interaction, lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry, Neuroscience, Gene, Astrocyte |
| الوصف: | Over the last decade, the importance of astrocyte-neuron communication in neuronal development and synaptic plasticity has become increasingly clear. Since neuron-astrocyte interactions represent highly dynamic and reciprocal processes, we hypothesized that many astrocyte genes may be regulated as a consequence of their interactions with maturing neurons. In order to identify such neuron-responsive astrocyte genes in vitro, we sought to establish an expedited technique for separation of neurons from co-cultured astrocytes. Our newly established method makes use of cold jet, which exploits different adhesion characteristics of subpopulations of cells (Jirsova et al., 1997), and is rapid, performed under ice-cold conditions and avoids protease-mediated isolation of astrocytes or time-consuming centrifugation, yielding intact astrocyte mRNA with approximately 90% of neuronal RNA removed. Using this purification method, we executed genome-wide profiling in which RNA derived from astrocyte-only cultures was compared with astrocyte RNA derived from differentiating neuron-astrocyte co-cultures. Data analysis determined that many astrocytic mRNAs and biological processes are regulated by neuronal interaction. Our results validate the cold jet as an efficient method to separate astrocytes from neurons in co-culture, and reveals that neurons induce robust gene-expression changes in co-cultured astrocytes. © 2014 Goudriaan, Camargo, Carney, Oliet, Smit and Verheijen. |
| تدمد: | 1662-5102 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::917fcd733dc0828ffff082aab883f280 https://pubmed.ncbi.nlm.nih.gov/24523672 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....917fcd733dc0828ffff082aab883f280 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16625102 |
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