Massively parallel digital transcriptional profiling of single cells

Grace Zheng(10X Genomics (United States)), Jessica M. Terry(10X Genomics (United States)), Phillip Belgrader(10X Genomics (United States)), Paul Ryvkin(10X Genomics (United States)), Zachary Bent(10X Genomics (United States)), Ryan J. Wilson(10X Genomics (United States)), Solongo B. Ziraldo(10X Genomics (United States)), Tobias D. Wheeler(10X Genomics (United States)), Geoff P. McDermott(10X Genomics (United States)), Junjie Zhu(10X Genomics (United States)), Mark Gregory(Fred Hutch Cancer Center), Joe Shuga(10X Genomics (United States)), Luz Montesclaros(10X Genomics (United States)), Jason G. Underwood(University of Washington), Donald A Masquelier(10X Genomics (United States)), Stefanie Y. Nishimura(10X Genomics (United States)), Michael Schnall-Levin(10X Genomics (United States)), Paul W. Wyatt(10X Genomics (United States)), Christopher M. Hindson(10X Genomics (United States)), Rajiv Bharadwaj(10X Genomics (United States)), Alexander Wong(10X Genomics (United States)), Kevin D. Ness(10X Genomics (United States)), Lan Beppu(Fred Hutch Cancer Center), H. Joachim Deeg(Fred Hutch Cancer Center), Christopher McFarland(Seattle Cancer Care Alliance), Keith R. Loeb(University of Washington), William J. Valente(University of Washington), Nolan G. Ericson(Fred Hutch Cancer Center), Emily A. Stevens(Fred Hutch Cancer Center), Jerald P. Radich(Fred Hutch Cancer Center), Tarjei S. Mikkelsen(10X Genomics (United States)), Benjamin J. Hindson(10X Genomics (United States)), Jason H. Bielas(University of Washington)
Nature Communications
January 16, 2017
Cited by 7,732Open Access
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Abstract

Characterizing the transcriptome of individual cells is fundamental to understanding complex biological systems. We describe a droplet-based system that enables 3' mRNA counting of tens of thousands of single cells per sample. Cell encapsulation, of up to 8 samples at a time, takes place in ∼6 min, with ∼50% cell capture efficiency. To demonstrate the system's technical performance, we collected transcriptome data from ∼250k single cells across 29 samples. We validated the sensitivity of the system and its ability to detect rare populations using cell lines and synthetic RNAs. We profiled 68k peripheral blood mononuclear cells to demonstrate the system's ability to characterize large immune populations. Finally, we used sequence variation in the transcriptome data to determine host and donor chimerism at single-cell resolution from bone marrow mononuclear cells isolated from transplant patients.


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