Journal Article

Fixed single-cell transcriptomic characterization of human radial glial diversity

Shehata, Soraya I, Yao, Zizhen, Doyle, Adele M, Ramanathan, Sharad, Mich, John K, Hodge, Rebecca D, Thomsen, Elliot R, Jang, Sumin, Shapovalova, Nadiya V, Nelson, Angelique M, Levi, Boaz P

2016

Overview

The Fixed and Recovered Intact Single-cell RNA (FRISCR) method enables robust RNA extraction and sequencing from fixed, stained and sorted single cells and allows unprecedented profiling of rare cell types, including two subpopulations of radial glial cells in the developing human cortex. 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.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 13/100 / 14/1 / 14/63 / 38/91 / 631/1647/2017 / 631/1647/514/1949 / 631/208/199 / 631/378/2571 / Bioinformatics / Biological Microscopy / Biological Techniques / Biomedical Engineering/Biotechnology / Brain - cytology / Cellular control mechanisms / Developmental biology / Gene expression / Genetic aspects / Genetic transcription / Humans / Life Sciences / Nervous system / Neuroglia / Neuroglia - cytology / Observations / Proteomics / Single-Cell Analysis / Transcriptome