The liver is an important organ for the regulation of whole-body metabolism, as well as for immunity. Kupffer cells (KCs) are specialized liver-resident macrophages and the major population of immune cells in the liver. These cells have been shown to play an important role for the regulation of liver homeostasis, and many studies have thus linked these cells to the development of various liver diseases. However, the complexity of macrophage populations and the lack of specific and exclusive markers have so far made it difficult to interpret results from many of these studies. Today, new technologies have emerged including next-generation sequencing allowing for more in depth investigation of multifaceted cell populations such as KCs. Here, we describe a protocol to isolate and prepare cDNA libraries for mRNA sequencing of murine liver macrophages. Using mRNA sequencing to study the gene expression of macrophages in the liver provides a great tool to study the various functions of these cells in the regulation of homeostasis and immunity.
Keywords: Bioinformatic analysis; Murine Kupffer cells (KCs); Next-generation sequencing; RNA extraction; mRNA; mRNA library preparation.