Gangliosides are a ubiquitous component of the membranes of mammalian cells that have been suggested to play important roles in various cell functions such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. However, the role that gangliosides play in the immune rejection response in xenotransplantation is not yet clearly understood. In this study, differential expression patterns of gangliosides in HEK293 (human embryonic kidney cells), PK15 (porcine kidney cells), NIH-kd (NIH-mini pig kidney cells, primary cultured) and the cortex, medulla and calyx of the NIH-mini pig kidney were investigated by high-performance thin-layer chromatography (HPTLC). The results revealed that HEK293, PK15 and NIH-kd contained GM3, GM2 and GD3 as major gangliosides. Moreover, GM3, which are the gangliosides of NIH-kd, were expressed at higher levels than HEK293 and PK15. Especially, GT1b were expressed in HEK293 and NIH-kd but not in PK15. Finally, GM1 and GD1a were expressed in NIH-kd, but not in HEK293 or PK15. These results suggest that differential expression patterns of gangliosides from HEK293, PK15 and NIH-kd are related to the immune rejection response in xenotransplantation.