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Nat Methods. 2016 Jan;13(1):87-93. doi: 10.1038/nmeth.3629. Epub 2015 Nov 16.

Fixed single-cell transcriptomic characterization of human radial glial diversity.

Author information

1
Allen Institute for Brain Science, Seattle, Washington, USA.
2
Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.
3
School of Engineering and Applied Science, Harvard University, Cambridge, Massachusetts, USA.

Abstract

The diverse progenitors that give rise to the human neocortex have been difficult to characterize because progenitors, particularly radial glia (RG), are rare and are defined by a combination of intracellular markers, position and morphology. To circumvent these problems, we developed Fixed and Recovered Intact Single-cell RNA (FRISCR), a method for profiling the transcriptomes of individual fixed, stained and sorted cells. Using FRISCR, we profiled primary human RG that constitute only 1% of the midgestation cortex and classified them as ventricular zone-enriched RG (vRG) that express ANXA1 and CRYAB, and outer subventricular zone-localized RG (oRG) that express HOPX. Our study identified vRG and oRG markers and molecular profiles, an essential step for understanding human neocortical progenitor development. FRISCR allows targeted single-cell profiling of any tissues that lack live-cell markers.

PMID:
26524239
PMCID:
PMC4869711
DOI:
10.1038/nmeth.3629
[Indexed for MEDLINE]
Free PMC Article

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