Protocol for isolation of primary hepatic lymphatic endothelial cells from rat liver

Here, we present a protocol for isolating and purifying primary rat hepatic lymphatic endothelial cells (hLECs) from an enriched population of hepatic endothelial cells (hECs). We describe procedures for rat liver perfusion, portal vascular tree isolation, and Percoll density gradient centrifugation. We then detail magnetic depletion of CD45⁺ cells, followed by fluorescence-activated cell sorting to obtain highly purified hLECs. This workflow overcomes the low abundance, fragility, and overlapping marker expression of hLECs with liver sinusoidal endothelial cells. For complete details on the use and execution of this protocol, please refer to Sharma et al.¹.