Cell Dissociation of Tissues for Single-Cell Analysis in African Killifish

The single-cell RNA sequencing (scRNA-seq) is a powerful tool to characterize cell heterogeneity, trace cell lineages, and identify novel cell markers in development, regeneration, and diseases. Although distinct platforms have been developed to conducting such analysis, the dissociation of tissues or organs to obtain the single-cell solution is an initial key step shared by all platforms. As an emerging model system, the single-cell RNA-seq has not yet been extensively used in African killifish compared with traditional model species. One of the challenges that hamper the application of this technology in African killifish is the lack of reliable protocols for producing a high-quality single-cell suspension. Here, we review tissue dissociation approaches that have been utilized in different systems. Using the regenerating fin blastema in African killifish as an example, we provide a reproducible and convenient protocol for the cell dissociation of the caudal fin tissue. Our protocol can be applied to other tissues in the African killifish with modifications for single-cell isolation.