RESUMO
It is now a routine to carry out single-cell RNA-Seq to define the gene expression patterns of thousands of cells, thereby revolutionizing many areas of research. Projects are underway to use these techniques to create an atlas of the expressed genes in all cell types of the human body. Here we describe cold-active protease methods for single-cell dissociation of organs and tissues that better preserve the in vivo gene expression patterns.
Assuntos
Análise de Célula Única/métodos , Perfilação da Expressão Gênica/métodos , Humanos , Análise de Sequência de RNA/métodosRESUMO
The heterogeneity and complexity of nonlymphoid tissues has become a major obstacle for the study of immune populations. For this reason, the generation of highly reproducible protocols that allow the analysis of immune cells in these tissues has become crucial for clinical and preclinical research. Here we describe an optimized method that allows the obtention of single-cell suspensions from the skin and lungs to analyze and quantify populations of tissue-resident memory CD8+ T cells by multi-parametric flow cytometry.