Subcutaneous adipocytes from obese hyperinsulinemic women with polycystic ovary syndrome exhibit normal insulin sensitivity but reduced maximal insulin responsiveness

Lystedt, Erika; Westergren, Hanna; Brynhildsen, Jan; Lindh-Åstrand, Lotta; Gustavsson, Johanna; Nyström, Fredrik; Hammar, Mats; Strålfors, Peter

doi:10.1530/eje.1.02027

Cited by 15 publications

(10 citation statements)

References 22 publications

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“…Rosenbaum et al reported a decrease in maximal insulin-stimulated glucose transport in adipocytes of women with PCOS [20], a defect that was not, however, confirmed by Ciaraldi et al [13]. Moreover, Lystedt et al found a normal insulin dose-response curve along with diminished maximal insulin-stimulated glucose transport in subcutaneous adipocytes of obese PCOS women [21]. Ciaraldi et al also showed decreased maximal insulin-stimulated autophosphorylation with no apparent defect in receptor kinase activity [13].…”

Section: Peripheral Tissuesmentioning

confidence: 98%

Defects in insulin signaling pathways in ovarian steroidogenesis and other tissues in polycystic ovary syndrome (PCOS)

Diamanti-Kandarakis

Argyrakopoulou

Economou

et al. 2008

The Journal of Steroid Biochemistry and Molecular Biology

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Section: Peripheral Tissuesmentioning

confidence: 98%

Defects in insulin signaling pathways in ovarian steroidogenesis and other tissues in polycystic ovary syndrome (PCOS)

Diamanti-Kandarakis

Argyrakopoulou

Economou

et al. 2008

The Journal of Steroid Biochemistry and Molecular Biology

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“…The differentiated adipocytes display phenotypic characteristics of genuine adipocytes, that is, freshly isolated ones. Specially, they respond to physiologically relevant concentrations of hormones, including insulin and β-adrenergic agonists (Fried et al, 2010; Lee & Fried, 2012; Lee, Wu, & Fried, 2012; Lystedt et al, 2005; Zierath et al, 1998). Further, ASCs are also useful for assessing donor- and origin-dependent effects (depot, sex, age, obesity, etc.)…”

Section: Introductionmentioning

confidence: 99%

Optimal Protocol for the Differentiation and Metabolic Analysis of Human Adipose Stromal Cells

Lee

Fried

2014

Methods in Enzymology

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Obesity is reaching epidemic proportions so there is growing interest in the mechanisms that regulates adipose tissue development and function. Although murine adipose cell lines are useful for many mechanistic studies, primary human adipose stromal cells (ASCs), which can be isolated from distinct adipose depots and cultured in vitro, have clear translational relevance. We describe the methods to isolate, culture, and differentiate human ASCs to adipocytes that respond to physiologically relevant hormones, such as insulin and β-adrenergic agonists. We also describe methods for assaying hormonal effects on glucose transport and lipolysis.

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“…Although several human adipose cell lines are available, including Simpson‐Golabi‐Behmel syndrome (SGBS) and Human multipotent adipose‐derived (hMAD) , primary preadipocytes isolated from adipose tissues can be differentiated into adipocytes in vitro and are useful for studies of donor or depot‐dependent effects . In addition, newly differentiated human adipocytes in culture respond to physiologically relevant concentrations of hormones, insulin, and beta‐adrenergic agonists . Thus, primary cultures of human preadipocytes and adipocytes are valuable tools for the studies of adipocyte development and function and their use is expected to increase.…”

Section: Introductionmentioning

confidence: 99%

Prolonged efficiency of siRNA‐mediated gene silencing in primary cultures of human preadipocytes and adipocytes

Lee

Pickering

Puri

2013

Obesity

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ObjectivePrimary human preadipocytes and differentiated adipocytes in culture are valuable cell culture systems to study adipogenesis and adipose function in relation to human adipose biology. To use these systems for mechanistic studies, we studied siRNA-mediated knockdown of genes for its effectiveness.Design and MethodsMethods were developed to effectively deliver siRNA to for gene silencing in primary preadipocytes isolated from human subcutaneous adipose tissue and newly-differentiated adipocytes. Expression level of genes and proteins was measured using quantitative RT-PCR and western blotting. Lipid droplet morphology was observed using microscopy and glycerol release was quantified as a measure of lipolysis.ResultssiRNA-mediated knockdown of genes in primary human preadipocytes resulted in prolonged silencing effects, suppressing genes throughout the process of their differentiation. In newly differentiated adipocytes, siRNA-mediated gene knockdown allowed proteins to stay depleted for at least 5 days. It was possible to re-express a protein after its siRNA-mediated depletion. Importantly, siRNA transfected human adipocytes remained metabolically active, responding to β-adrenergic stimulation to increase lipolysis.ConclusionsOur study describes the methods of gene silencing in primary cultures of human preadipocytes and adipocytes and their prolonged effectiveness.

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Subcutaneous adipocytes from obese hyperinsulinemic women with polycystic ovary syndrome exhibit normal insulin sensitivity but reduced maximal insulin responsiveness

Cited by 15 publications

References 22 publications

Defects in insulin signaling pathways in ovarian steroidogenesis and other tissues in polycystic ovary syndrome (PCOS)

Defects in insulin signaling pathways in ovarian steroidogenesis and other tissues in polycystic ovary syndrome (PCOS)

Optimal Protocol for the Differentiation and Metabolic Analysis of Human Adipose Stromal Cells

Prolonged efficiency of siRNA‐mediated gene silencing in primary cultures of human preadipocytes and adipocytes

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