Preparation of Single Pineal Cells

The isolation of single cells from the pineal gland plays an essential role in understanding the complex nature of such processes as differentiation, metabolism, and cell-cell communication within the pineal gland. This procedure is the portal to single-cell RNA sequencing, which produces the transcriptome of individual cells. As such, single-cell RNA sequencing is critical to the continued development of knowledge of the pineal cell physiology. This chapter describes a simple procedure for isolating individual cells. Starting with the incubation of whole tissue in an enzyme preparation, which dissociates the pineal gland into small pieces, it continues with gentle trituration and then isolation of single cells through filtration. The procedure takes less than 2 h.