A Rapid, Calcium-independent Mojosort &[trade] Dead Cell Removal Kit for Improved Live Cell Enrichment and Downstream Application

Abstract

Abstract High sample cell viability is crucial for obtaining high-quality data for various downstream applications. Commonly used methods include density centrifugation, flow cytometry sorting, and Annexin V-based magnetic cell separation. However, above entioned methods have several limitations that ultimately leads to fewer live cell recovery. Here we present MojoSort™ dead cell removal kit, which relies on Ca2+-independent, Phoshatidylserine (PS)-binding for dead and apoptotic cell removal. Unlike Annexin V that requires specialized, high Ca2+-containing buffers that can potentially induce cytotoxicity, MojoSort™ dead cell removal kit can be performed in the buffer with or without Ca2+ that also contain chelators such as EDTA. This enables 20% improved live cell recovery compared to other commercially available, Annexin V-based magnetic cell separation kits, while demonstrating 90–95% live peripheral blood mononuclear cell (PBMC) purity post-enrichment from 50% starting viability. Single-cell cellular Indexing of transcriptomes and epitopes by sequencing (CITE-seq) demonstrated effective removal of a distinct dead cell cluster comprised of cells with high mitochondrial gene read counts, while preserving all common PBMC lineages and surface marker expression levels. In summary, MojoSort™ dead cell removal provides a rapid, effective live cell enrichment option that overcomes shortcomings associated with other routinely used methods for downstream applications that require high sample viability for success.