Protocol for isolating and purifying murine liver sinusoidal endothelial cells for in vitro culture and functional assays

Liver sinusoidal endothelial cells (LSECs) line the liver sinusoids and play a crucial role in liver function. Isolating LSECs is beneficial for their functional evaluation in vitro. Here, we provide a protocol for obtaining purified LSECs from mice via gradient centrifugation and magnetic cell sorting (MACS), yielding cells suitable for culture and downstream analyses. We describe steps for culturing the purified LSECs and demonstrate their evaluation by flow cytometry. For complete details on the use and execution of this protocol, please refer to Papaioannou et al.1.