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

Nature methods (2015-11-03)
Elliot R Thomsen, John K Mich, Zizhen Yao, Rebecca D Hodge, Adele M Doyle, Sumin Jang, Soraya I Shehata, Angelique M Nelson, Nadiya V Shapovalova, Boaz P Levi, Sharad Ramanathan
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.

MATERIALS
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Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, fatty acid free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
Sigma-Aldrich
DNQX, ≥98% (TLC)
Sigma-Aldrich
Triton X-100, laboratory grade