Culture of Isolated Human Adipocytes and Isolated Adipose Tissue

Abstract

Adipose tissue (AT) is no longer considered merely as insulation or padding for human organs. It is an endocrine organ in its own right, which includes composite cells with the ability to differentiate into multiple cell lines. In fact, there is increasing evidence to support the theory that the causation of obesity and its associated metabolic disorders originate at the cellular or tissue level. Adipocyte dysfunction and chronic inflammatory states are able to modulate triglyceride storage and mobilization directly through cytokine and adipokine release. Significant variability exists between adipocyte isolation and culture techniques which subsequently can impact experimental results. We aim to explain the importance of controlling these variables, to assist tailoring methodological choice towards the investigational outcomes, and modifications of the techniques used accordingly. The techniques described in this chapter yield cell and adipose tissue which can be utilised in many different ways, including adipose tissue stem cells for differentiation, DNA analysis, RT-PCR, immunohistochemistry, lipolysis, glucose uptake, and LPL activity.

Potential for in vitro mesoderm differentiation of Wharton's jelly cells from ovine umbilical cord isolated in different culture media

“…Low glucose DMEM and M199 allowed cells to respond better to this induction, with a discreet but visible superiority when compared to MEM (Fig.3B,C). These data corroborate with the methodology proposed by Carswell et al (2012), which indicate both DMEM and M199 as the ideal media for isolation and culture of adipocytes. Janderová et al (2003) report a superior differentiation in human mesenchymal stem cells into adipocytes provided by M199 added with adipogenic factors, when compared to high glucose DMEM.…”

Addition of Exercise Increases Plasma Adiponectin and Release from Adipose Tissue

“…Our study suggests that changes in adiponectin release from gluteal fat were in the same direction as that from abdominal fat, but of less magnitude. We recognize that the adiponectin release method we used is limited to the tissue collected from the biopsy, and Bovine Serum Albumin that was used in the media may influence experimental outcome as it may contain growth factors(14). This method quantifies the amount of adiponectin released from the adipose tissue during the incubation period, which represents synthesis by adipocytes and other cells in the tissue; however, whether cells other than adipocytes contributes to circulating adiponectin in humans is still unanswered(19).…”

GPS2/KDM4A Pioneering Activity Regulates Promoter-Specific Recruitment of PPARγ

“…Lipolysis assays were performed in mice primary adipocytes, isolated as described (Carswell et al 2012), or in mature 3T3-L1 cells after 6–8 day of induced differentiation. 3T3-L1 cells were grown, transfected and induced to differentiate as previously described (Cardamone et al 2012).…”