Understanding the function of fat metabolism during differentiation of human preadipocytes to fully developed fat tissue has been the aim of various studies in the past decades. Due to the lack of suitable human cell culture lines, experimental research predominantly focused on rodent models and nonhuman cell culture systems. Here, we demonstrate that a human preadipocyte cell line SGBS is well suited to examine differential expression of the Acyl-CoA binding protein (ACBP) during adipogenesis. The Acbp gene expresses various alternative high- and low-abundant transcript variants encoding ACBP protein isoforms, which play a central role in fat metabolism. Whereas the low-abundant transcript Acbp-1G is downregulated during SGBS adipogenesis, the high-abundant and well established transcripts Acbp-1A (1) and -1B are moderately (2-4-fold) upregulated. In contrast, the alternative high-abundant transcript Acbp-1C is strongly (29-fold) upregulated at mRNA and protein level indicating that particularly ACBP-1C functions in lipogenic processes during fat cell differentiation in humans.