从 COVID-19 患者外周血免疫细胞中进行样本处理和单细胞 RNA 测序。
Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients.
机构信息
Women's Guild Lung Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Division of Pulmonary and Critical Care Medicine, Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
出版信息
STAR Protoc. 2021 Jun 18;2(2):100582. doi: 10.1016/j.xpro.2021.100582. Epub 2021 May 12.
Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows in-depth assessment of transcriptional changes in immune cells of patients with COVID-19. However, collecting, processing, and analyzing samples from patients with COVID-19 pose many challenges because blood samples may contain infectious virus, identification of immune cell subtypes can be difficult, and biological interpretation of analytical results is complex. To address these issues, we describe a protocol for sample processing, sorting, methanol fixation, and scRNA-seq analysis of PBMCs from frozen buffy coat samples from patients with COVID-19. For complete details on the use and execution of this protocol, please refer to (Yao et al., 2021).
单细胞 RNA 测序 (scRNA-seq) 可深入评估 COVID-19 患者免疫细胞的转录变化。然而，收集、处理和分析 COVID-19 患者的样本存在诸多挑战，因为血液样本可能含有传染性病毒，免疫细胞亚型的鉴定可能较为困难，并且分析结果的生物学解释也较为复杂。为了解决这些问题，我们描述了一种从 COVID-19 患者冷冻 buffy coat 样本的 PBMC 中进行样本处理、分选、甲醇固定和 scRNA-seq 分析的方案。有关此方案使用和执行的完整详细信息，请参见 (Yao 等人，2021)。
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