Assessing similarity to primary tissue and cortical layer identity in induced pluripotent stem cell-derived cortical neurons through single-cell transcriptomics

Induced pluripotent stem cell (iPSC)-derived cortical neurons potentially present a powerful new model to understand corticogenesis and neurological disease. Previous work has established that differentiation protocols can produce cortical neurons, but little has been done to characterize these at c...

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Veröffentlicht in:Human molecular genetics Jg. 25; H. 5; S. 989
Hauptverfasser: Handel, Adam E, Chintawar, Satyan, Lalic, Tatjana, Whiteley, Emma, Vowles, Jane, Giustacchini, Alice, Argoud, Karene, Sopp, Paul, Nakanishi, Mahito, Bowden, Rory, Cowley, Sally, Newey, Sarah, Akerman, Colin, Ponting, Chris P, Cader, M Zameel
Format: Journal Article
Sprache:Englisch
Veröffentlicht: England 01.03.2016
Schlagworte:
Adult
Aged
Biomarkers - metabolism
Cell Differentiation
Cell Line
Cerebral Cortex - cytology
Cerebral Cortex - metabolism
Female
Fetus
Fibroblasts - cytology
Fibroblasts - metabolism
Gene Expression Profiling
Gene Expression Regulation, Developmental
Humans
Induced Pluripotent Stem Cells - cytology
Induced Pluripotent Stem Cells - metabolism
Nerve Tissue Proteins - genetics
Nerve Tissue Proteins - metabolism
Neurogenesis - genetics
Neurons - cytology
Neurons - metabolism
Real-Time Polymerase Chain Reaction
Receptors, Glutamate - genetics
Receptors, Glutamate - metabolism
Sequence Analysis, RNA
Single-Cell Analysis
Transcriptome
ISSN:1460-2083, 1460-2083
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Zusammenfassung:Induced pluripotent stem cell (iPSC)-derived cortical neurons potentially present a powerful new model to understand corticogenesis and neurological disease. Previous work has established that differentiation protocols can produce cortical neurons, but little has been done to characterize these at cellular resolution. In particular, it is unclear to what extent in vitro two-dimensional, relatively disordered culture conditions recapitulate the development of in vivo cortical layer identity. Single-cell multiplex reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) was used to interrogate the expression of genes previously implicated in cortical layer or phenotypic identity in individual cells. Totally, 93.6% of single cells derived from iPSCs expressed genes indicative of neuronal identity. High proportions of single neurons derived from iPSCs expressed glutamatergic receptors and synaptic genes. And, 68.4% of iPSC-derived neurons expressing at least one layer marker could be assigned to a laminar identity using canonical cortical layer marker genes. We compared single-cell RNA-seq of our iPSC-derived neurons to available single-cell RNA-seq data from human fetal and adult brain and found that iPSC-derived cortical neurons closely resembled primary fetal brain cells. Unexpectedly, a subpopulation of iPSC-derived neurons co-expressed canonical fetal deep and upper cortical layer markers. However, this appeared to be concordant with data from primary cells. Our results therefore provide reassurance that iPSC-derived cortical neurons are highly similar to primary cortical neurons at the level of single cells but suggest that current layer markers, although effective, may not be able to disambiguate cortical layer identity in all cells.
Bibliographie:ObjectType-Article-1
SourceType-Scholarly Journals-1
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content type line 23
ISSN:1460-2083
1460-2083
DOI:10.1093/hmg/ddv637