液滴微流控单细胞转录组检测技术的建立和优化

doi:10.3969/j.issn.1004-616x.2019.01.002

Carcinogenesis, Teratogenesis & Mutagenesis ›› 2019, Vol. 31 ›› Issue (1): 9-14,21.doi: 10.3969/j.issn.1004-616x.2019.01.002

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Establishment and optimization of droplet microfluidic single-cell transcriptome detection technology

ZHOU Lulin¹, YANG Runkun², ZHANG Wen³, FENG Lin¹, XIAO Ting¹, CHENG Shujun¹, ZHANG Kaitai¹, ZHANG Lei⁴

1. State Key Laboratory of Molecular Oncology, Beijing Key Laboratory for Carcinogenesis and Cancer Prevention, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021;
2. Department of Colorectal Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin 150086;
3. Department of Immunology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021;
4. Department of Endoscope, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China

Received:2018-11-15 Revised:2018-12-29 Online:2019-01-31 Published:2019-01-31

Abstract

Abstract: OBJECTIVE:To establish and to validate a droplet microfluidic single-cell transcriptome detection platform for cancer research. METHODS:Using a droplet microfluidic system,a single cell and a single barcoded microparticle were wrapped together in an oil-water emulsion droplet. The mixed human acute promyelocytic leukemia cell line HL-60 and the mouse melanoma cell line B16-F10 were used as simulative sample to evaluate the technical indexes of the system. RESULTS:When the oil and the water phase flow rates were 200 and 30 μL/min,respectively,and the cell and microparticle concentrations were at 250 and 400/μL,the optimum cell labeling rate was reached at (11.87±3.75)%. With a simulative sample which contained 75 thousand cells and the theoretical cell labeling rate of 11%,the transcriptome data of 7 535 cells was obtained,among which there were 70 mixed human and mouse transcriptome cells accounting for only 0.9% of all cells. Transcriptome differences were used to distinguish the two types of cells and to screen out the specific expression markers among them. CONLUSION:In this study,a droplet microfluidic single-cell transcriptome detection platform was initiated and validated. The platform meets the requirements of oncology laboratory and shows extensive potential for tumor research.

Key words: single-cell sequencing, microfluidic, transcriptome, tumor cells

CLC Number:

R-331

ZHOU Lulin, YANG Runkun, ZHANG Wen, FENG Lin, XIAO Ting, CHENG Shujun, ZHANG Kaitai, ZHANG Lei. Establishment and optimization of droplet microfluidic single-cell transcriptome detection technology[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 9-14,21.

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Establishment and optimization of droplet microfluidic single-cell transcriptome detection technology

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