Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/93695
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dc.contributor.authorSu, Colin T. E.en
dc.contributor.authorYoon, Su-Inen
dc.contributor.authorMarcy, Guillaumeen
dc.contributor.authorChin, Eunice W. M.en
dc.contributor.authorAugustine, George Jamesen
dc.contributor.authorGoh, Eyleen Lay Keowen
dc.date.accessioned2015-07-15T08:00:42Zen
dc.date.accessioned2019-12-06T18:43:49Z-
dc.date.available2015-07-15T08:00:42Zen
dc.date.available2019-12-06T18:43:49Z-
dc.date.copyright2015en
dc.date.issued2015en
dc.identifier.citationSu, C. T. E., Yoon, S.-I., Marcy, G., Chin, E. W. M., Augustine, G. J., & Goh, E. L. K. (2015). An optogenetic approach for assessing formation of neuronal connections in a co-culture system. Journal of Visualized Experiments, 96, e52408.en
dc.identifier.issn1940-087Xen
dc.identifier.urihttps://hdl.handle.net/10356/93695-
dc.description.abstractHere we describe a protocol to generate a co-culture consisting of 2 different neuronal populations. Induced pluripotent stem cells (iPSCs) are reprogrammed from human fibroblasts using episomal vectors. Colonies of iPSCs can be observed 30 days after initiation of fibroblast reprogramming. Pluripotent colonies are manually picked and grown in neural induction medium to permit differentiation into neural progenitor cells (NPCs). iPSCs rapidly convert into neuroepithelial cells within 1 week and retain the capability to self-renew when maintained at a high culture density. Primary mouse NPCs are differentiated into astrocytes by exposure to a serum-containing medium for 7 days and form a monolayer upon which embryonic day 18 (E18) rat cortical neurons (transfected with channelrhodopsin-2 (ChR2)) are added. Human NPCs tagged with the fluorescent protein, tandem dimer Tomato (tdTomato), are then seeded onto the astrocyte/cortical neuron culture the following day and allowed to differentiate for 28 to 35 days. We demonstrate that this system forms synaptic connections between iPSC-derived neurons and cortical neurons, evident from an increase in the frequency of synaptic currents upon photostimulation of the cortical neurons. This co-culture system provides a novel platform for evaluating the ability of iPSC-derived neurons to create synaptic connections with other neuronal populations.en
dc.language.isoenen
dc.relation.ispartofseriesJournal of visualized experimentsen
dc.rights© 2015 Creative Commons Attribution-NonCommercial License. This paper was published in Journal of Visualized Experiments and is made available as an electronic reprint (preprint) with permission of Creative Commons Attribution-NonCommercial License. The published version is available at: [http://dx.doi.org/10.3791/52408]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.en
dc.subjectDRNTU::Science::Biological sciences::Human anatomy and physiology::Neurobiologyen
dc.titleAn optogenetic approach for assessing formation of neuronal connections in a co-culture systemen
dc.typeJournal Articleen
dc.contributor.schoolLee Kong Chian School of Medicine (LKCMedicine)en
dc.identifier.doi10.3791/52408en
dc.description.versionPublished versionen
dc.identifier.pmid25742527-
item.grantfulltextopen-
item.fulltextWith Fulltext-
Appears in Collections:LKCMedicine Journal Articles

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