Abstract: Objective To establish a method for primary culture of human preadipocyte as the seed cells for adipose tissue engineering.Methods Fibroblast-like cells were cultured from adult human abdominal adipose tissue. The morphological changes of the cultured cells were observed and the growth curve drawn. The intracytoplasmic lipid of the cultured cells was determined using oil red O staining.Results The cultured fibroblast-like cells showed highly homogeneous appearance with active proliferation, and could differentiate into mature adipocytes. Oil red O staining and morphological and biological observation verified these cells as preadipocytes with accumulation of lipid droplets in the cytoplasm. Extraction of the intracytoplasmic lipid stained with oil red O suggested a rapid increase in the intracytoplasmic lipid content on the 9th day of inoculation, which peaked on day 18.Conclusions Preadipocytes are present in mature human adipose tissue and possess the potential to proliferate and differentiate into mature adipocytes. These homogeneous preadipocytes with good proliferating activity can be harvested through primary cell culture.