Isolation of planarian viable cells using fluorescence-activated cell sorting for advancing single-cell transcriptome analysis.

ABSTRACT: Preparing viable single cells is critical for conducting single-cell RNA sequencing (scRNA-seq) because the presence of ambient RNA from dead or damaged cells can interfere with data analysis. Here, we developed a method for isolating viable single cells from adult planarian bodies using fluorescence-activated cell sorting (FACS). This method was then applied to both adult pluripotent stem cells (aPSCs) and differentiating/differentiated cells. Initially, we employed a violet instead of ultraviolet (UV) laser to excite Hoechst 33342 to reduce cellular damage. After optimization of cell staining conditions and FACS compensation, we generated FACS profiles similar to those created using a previous method that employed a UV laser. Despite successfully obtaining high-quality RNA sequencing dat

SUBMITTER: Kuroki Y 

PROVIDER: S-EPMC11448005 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature