Massively parallel digital transcriptional profiling of single cells
Массивно-параллельное цифровое транскриптомное профилирование отдельных клеток
2017-01-16
SCID: 54.1/fg5rh3kv
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3' mRNA countingdroplet-based systemhost-donor chimerismperipheral blood mononuclear cellssingle-cell transcriptomics
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Abstract (AI)
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.
Key Findings
1
A droplet-based system enables massively parallel 3′ mRNA counting of tens of thousands of single cells per sample.
2
Profiling 68,000 peripheral blood mononuclear cells showed that the system can characterize large immune-cell populations.
3
The system generated transcriptome profiles for approximately 250,000 single cells across 29 samples and demonstrated sensitivity for detecting rare populations.
4
Transcript sequence variation enabled single-cell resolution of host–donor chimerism in bone marrow cells from transplant patients.
5
Up to eight samples can be encapsulated simultaneously in approximately 6 minutes, with approximately 50% cell-capture efficiency.
Research Object
Droplet-based single-cell 3' mRNA counting system for massively parallel transcriptional profiling
Research Subject
massively parallel 3′ mRNA transcriptome profiling and characterization of cell populations, including rare populations and host–donor chimerism, at single-cell resolution
Publication Details
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2017-01-16
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