Preparation of mouse pancreatic tumor for single-cell RNA sequencing and analysis of the data

被引:1
|
作者
Surumbayeva, Aizhan [1 ]
Kotliar, Michael [2 ,5 ]
Gabitova-Cornell, Linara [1 ]
Kartashov, Andrey [2 ,5 ,6 ]
Peri, Suraj [7 ,8 ]
Salomonis, Nathan [4 ,5 ]
Barski, Artem [1 ,3 ,5 ,6 ]
Astsaturov, Igor [1 ,8 ]
机构
[1] Fox Chase Canc Ctr, Marvin & Concetta Greenberg Pancreat Canc Inst, Philadelphia, PA 19111 USA
[2] Univ Cincinnati, Div Allergy & Immunol, Cincinnati, OH 45221 USA
[3] Univ Cincinnati, Div Human Genet, Cincinnati, OH 45221 USA
[4] Univ Cincinnati, Div Biomed Informat, Cincinnati, OH 45221 USA
[5] Univ Cincinnati, Cincinnati Childrens Hosp Med Ctr, Dept Pediat, Cincinnati, OH 45221 USA
[6] Datirium LLC, Cincinnati, OH 45226 USA
[7] Fox Chase Canc Ctr, Biostat & Bioinformat Facil, Philadelphia, PA 19111 USA
[8] Fox Chase Canc Ctr, Canc Biol Program, Philadelphia, PA 19111 USA
来源
STAR PROTOCOLS | 2021年 / 2卷 / 04期
关键词
Bioinformatics; Cancer; Cell Biology; Cell isolation; Molecular Biology; RNAseq; Sequence analysis; Single Cell;
D O I
10.1016/j.xpro.2021.100989
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Preparation of single-cell suspension from primary tumor tissue can provide a valuable resource for functional, genetic, proteomic, and tumor microenvironment studies. Here, we describe an effective protocol for mouse pancreatic tumor dissociation with further processing of tumor suspension for single-cell RNA sequencing analysis of cellular populations. We further provide an outline of the bioinformatics processing of the data and clustering of heterogeneous cellular populations comprising pancreatic tumors using Common Workflow Language (CWL) pipelines within user-friendly Scientific Data Analysis Platform (https://SciDAP.com). For complete details on the use and execution of this protocol, please refer to Gabitova-Cornell et al. (2020).
引用
收藏
页数:24
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