Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and

被引:2
作者
Jia, Jie [1 ,2 ,3 ]
Zhao, Yu [1 ,2 ,3 ]
Wang, Jian-Hua [4 ]
Kuang, Yi-Qun [1 ,2 ,3 ]
机构
[1] Kunming Med Univ, Affiliated Hosp 1, NHC Key Lab Drug Addict Med, Kunming 650032, Yunnan, Peoples R China
[2] Kunming Med Univ, Yunnan Key Lab Stem Cell & Regenerat Med, Kunming 650032, Yunnan, Peoples R China
[3] Kunming Med Univ, Affiliated Hosp 1, Sci Res Lab Ctr, Kunming 650032, Yunnan, Peoples R China
[4] Chinese Acad Sci, Guangzhou Inst Biomed & Hlth, Guangzhou 510530, Guangdong, Peoples R China
来源
STAR PROTOCOLS | 2023年 / 4卷 / 02期
基金
中国国家自然科学基金;
关键词
Bioinformatics; Cell Biology; Cell Isolation; Cell-based Assays; Immunology; Molecular Biology; RNAseq; Sequence Analysis; Single Cell;
D O I
10.1016/j.xpro.2023.102222
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows the dissection of transcriptional changes in immune cells with HIV infection. Here, we isolate PBMCs from HIV-infected individuals. After counting the cell number and verifying cell viability, we perform scRNA-seq for PBMCs on the 103 Genomics protocol and the Illumina NovaSeq 6000 sequencing platform. Furthermore, we analyze the function and cellular trajectories of B cell subsets and B cell receptor (BCR) repertoire after filtering raw sequences data and normalizing gene expression.For complete details on the use and execution of this protocol, please refer to Jia et al. (2022).
引用
收藏
页数:22
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共 14 条
  • [11] An optimized protocol to identify keratinocyte subpopulations in vitro by single-cell RNA sequencing analysis
    Siriwach, Ratklao
    Ngo, Anh Quynh
    Narumiya, Shuh
    Thumkeo, Dean
    [J]. STAR PROTOCOLS, 2022, 3 (04):
  • [12] Comprehensive Integration of Single-Cell Data
    Stuart, Tim
    Butler, Andrew
    Hoffman, Paul
    Hafemeister, Christoph
    Papalexi, Efthymia
    Mauck, William M., III
    Hao, Yuhan
    Stoeckius, Marlon
    Smibert, Peter
    Satija, Rahul
    [J]. CELL, 2019, 177 (07) : 1888 - +
  • [13] Preparation of mouse pancreatic tumor for single-cell RNA sequencing and analysis of the data
    Surumbayeva, Aizhan
    Kotliar, Michael
    Gabitova-Cornell, Linara
    Kartashov, Andrey
    Peri, Suraj
    Salomonis, Nathan
    Barski, Artem
    Astsaturov, Igor
    [J]. STAR PROTOCOLS, 2021, 2 (04):
  • [14] The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells
    Trapnell, Cole
    Cacchiarelli, Davide
    Grimsby, Jonna
    Pokharel, Prapti
    Li, Shuqiang
    Morse, Michael
    Lennon, Niall J.
    Livak, Kenneth J.
    Mikkelsen, Tarjei S.
    Rinn, John L.
    [J]. NATURE BIOTECHNOLOGY, 2014, 32 (04) : 381 - U251