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Massively parallel digital transcriptional profiling of single cells

Nature Communications · 2017 · Vol. 8(1) · pp. 14049–14049
Grace ZhengJessica M. TerryPhillip BelgraderPaul RyvkinZachary BentRyan J. WilsonSolongo B. ZiraldoTobias D. WheelerGeoff P. McDermottJunjie ZhuMark GregoryJoe ShugaLuz MontesclarosJason G. UnderwoodDonald A MasquelierStefanie Y. NishimuraMichael Schnall-LevinPaul W. WyattChristopher M. HindsonRajiv BharadwajAlexander WongKevin D. NessLan BeppuH. Joachim DeegChristopher McFarlandKeith R. LoebWilliam J. ValenteNolan G. EricsonEmily A. StevensJerald P. RadichTarjei S. MikkelsenBenjamin J. HindsonJason H. Bielas

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.

Single-cell and spatial transcriptomicsCancer Genomics and DiagnosticsGene Regulatory Network AnalysisTranscriptomePeripheral blood mononuclear cellComputational biologySingle-cell analysisBiologyGene expression profilingCellCell biologyGeneGene expression

MeSH terms

Cell LineFemaleHumansLeukocytes, MononuclearMaleRNA, MessengerSingle-Cell AnalysisTranscriptome

Funding

  • U.S. Department of Defense
  • Ellison Medical Foundation
  • Canary Foundation
  • National Cancer Institute
  • National Institute of Environmental Health Sciences
  • Congressionally Directed Medical Research Programs
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