Massively parallel digital transcriptional profiling of single cells.
Massively parallel digital transcriptional profiling of single cells.
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DOI：
10.1038/ncomms14049
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发表时间：
2017-01-16
影响因子：
16.6
通讯作者：
Bielas JH
中科院分区：
文献类型：
--
作者：
Zheng GX;Terry JM;Belgrader P;Ryvkin P;Bent ZW;Wilson R;Ziraldo SB;Wheeler TD;McDermott GP;Zhu J;Gregory MT;Shuga J;Montesclaros L;Underwood JG;Masquelier DA;Nishimura SY;Schnall-Levin M;Wyatt PW;Hindson CM;Bharadwaj R;Wong A;Ness KD;Beppu LW;Deeg HJ;McFarland C;Loeb KR;Valente WJ;Ericson NG;Stevens EA;Radich JP;Mikkelsen TS;Hindson BJ;Bielas JH
表征单个细胞的转录组对于理解复杂的生物系统至关重要。我们描述了一种基于液滴的系统，该系统能够对每个样本中数以万计的单个细胞进行3′ mRNA计数。一次可对多达8个样本进行细胞封装，耗时约6分钟，细胞捕获效率约为50%。为了证明该系统的技术性能，我们从29个样本中的约25万个单个细胞收集了转录组数据。我们验证了该系统的敏感性以及其使用细胞系和合成RNA检测稀有细胞群的能力。我们对6.8万个外周血单个核细胞进行了分析，以证明该系统表征大型免疫细胞群的能力。最后，我们利用转录组数据中的序列变异，从移植患者分离的骨髓单个核细胞中以单细胞分辨率确定宿主和供体的嵌合情况。
单细胞基因表达分析具有挑战性。这项工作描述了一种新的基于液滴的单细胞RNA - seq平台，该平台能够在数分钟内处理8个独立样本中的数以万计的细胞，并证明了移植患者中的细胞亚型以及宿主 - 供体嵌合情况。
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 gene expression analysis is challenging. This work describes a new droplet-based single cell RNA-seq platform capable of processing tens of thousands of cells across 8 independent samples in minutes, and demonstrates cellular subtypes and host–donor chimerism in transplant patients.