Isolation of Mature Adipocytes from White Adipose Tissue and Gene Expression Studies by Real-Time Quantitative RT-PCR.

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

• Level A · Stronger Clinical Evidence
• Level B · Emerging clinical evidence with positive signals
• Level C · Early human research exploring benefits
• Level D · Scientific groundwork from lab and animal studies
• Emerging · Emerging topic under active research

Abstract (original English)

The study of adipose tissue and more specifically of adipocytes is considered pivotal for dissecting molecular mechanisms responsible for alterations in several organs and systems, including adipose tissue, not only in obesity but also in other diseases (hypertension, heart failure). Adipose tissue is a complex tissue composed of adipocytes and the stromal vascular fraction which includes a heterogeneous population of preadipocytes, blood cells, endothelial cells, and macrophages. In the present chapter, methods are detailed to generate purified mature adipocytes from white adipose tissue by using enzymatic digestion. Such methods should help laboratories to study the specific roles of adipocytes in different pathologies and are easily adaptable to different animal models. Moreover, as gene activity is controlled at both transcriptional and posttranscriptional levels, it is very important to determine the levels of messenger ribonucleic acid (mRNA) of genes of interest. This process involves the isolation of total RNA and subsequent analysis of the mRNA of interest by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Given the unique nature of adipose tissue and adipocytes (i.e., containing high amounts of lipid), we have set up a special RNA isolation technique in both white adipose tissue and isolated mature adipocytes from mice. In summary, is

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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