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Human Primary Preadipocytes and Differentiated Adipocytes

because humans and rodents have distinct genetic backgrounds and metabolic pathways, potential experimental pitfalls were inevitable. For example, compounds effective in activating the b3-adrenergic pathway in rodents failed to stimulate human adipocytes to burn more fat.3 Stratagene now offers Human Predifferentiated Adipocytes and Human Differentiated Adipocytes, so rodent systems can be bypassed for human systems in experiments involving adipocyte signal transduction pathways or gene regulation.

Human Preadipocyte Differentiation

When excess preadipocytes are recruited into the differentiated adipocyte, obesity results. Since from adolescence through adulthood humans continue to recruit cells to become adipocytes, it is of interest to study the environmental and humoral factors that initiate and maintain the differentiated phenotype of adipocytes. Human preadipocytes provide the tools to explore the most relevant factors that influence human obesity.

Stratagene recently developed the Adipocyte Differentiation Kit, which uses a simple and efficient method to differentiate human primary preadipocytes into mature, fully functional adipocytes. Undifferentiated preadipocytes are cultured in DMEM/F-10 (1:1, v/v) with 10% fetal bovine serum. The cells can be passaged for expansion, frozen for later reuse, or differentiated. The absence of staining with antisera to factor VIII and lack of vascular endothelial growth factor receptor mRNA, shown by Northern analysis, 4 demonstrated that no endothelial cells are present in the preadipocyte culture.

Earlier passage cells are more robust in both growth and differentiation potential; passages one to three are ideal. Stratagene uses only passage-two human preadipocytes for the Adipocyte Differentiation Kit. Typically, a production lot of cells is isogenic, and lots vary within the 10% to 20% range
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