PBMC fixation and processing for Chromium single-cell RNA sequencing.

ABSTRACT:

Background

Interest in single-cell transcriptomic analysis is growing rapidly, especially for profiling rare or heterogeneous populations of cells. In almost all reported works investigators have used live cells, which introduces cell stress during preparation and hinders complex study designs. Recent studies have indicated that cells fixed by denaturing fixative can be used in single-cell sequencing, however they did not usually work with most types of primary cells including immune cells.

Methods

The methanol-fixation and new processing method was introduced to preserve human peripheral blood mononuclear cells (PBMCs) for single-cell RNA sequencing (scRNA-Seq) analysis on 10× Chromium platform.

Results

When methanol fixation protocol was broken up into three steps:

SUBMITTER: Chen J 

PROVIDER: S-EPMC6050658 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature