The metabolism of isolated fat cells

Rodbell, Martin

doi:10.1002/cphy.cp050147

Cited by 180 publications

(177 citation statements)

References 2,275 publications

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“…All visible connective tissue, blood and other debris were removed. The adipocytes were then isolated using the collagenase digestion technique (Rodbell, 1964): samples were incubated at 39°C in 5 ml of M199 medium (Gibco 31150-022, Waltham, MA, USA) (pH: 7.0 to 7.4) containing 200 mg of bovine serum albumin (Sigma A7906, Sigma-Aldrich, Madrid, Spain) and 5 mg (SC AT) or 10 mg (LM) type 2 collagenase (Sigma C6885) in an incubator for 2 h. After digestion, the samples were filtered through a nylon sieve (850 μm mesh size). The adipocytes so isolated were recovered from the supernatant and examined on microscope slides.…”

Section: Methodsmentioning

confidence: 99%

Expression of genes involved in adipogenesis and lipid metabolism in subcutaneous adipose tissue and longissimus muscle in low-marbled Pirenaica beef cattle

Soret

Mendizábal

Arana

et al. 2016

animal

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The ability to accumulate intramuscular fat (IMF) is a highly variable characteristic in beef cattle. In breeds with a low tendency to accumulate IMF, this can lead to compromised meat quality because of the contribution of fat to such organoleptic attributes as juiciness and taste. This study considered adiposity and gene expression of some of the main markers involved in adipogenesis and lipid metabolism in the subcutaneous (SC) adipose tissue (AT) and the longissimus thoracis muscle (LM) and investigated differences in adipogenic regulation between the tissues during growth and fattening under different conditions. Pirenaica beef cattle were chosen for the study due to the breed's low tendency to accumulate IMF and the breed's regional importance. The young Pirenaica bulls used (n = 16) were allocated to four groups and slaughtered at 6, 12 and 18 months. From 12 months onwards the bulls slaughtered at 18 months were fed diets having different energy densities. Backfat thickness increased from 6 to 12 months (P < 0.05) but then was unchanged, while other fattening parameters such as percentage chemical fat and marbling did not vary. The adipose cell size distribution displayed a bimodal distribution for SC adipocytes and a unimodal distribution for IMF cells, suggestive of tissue-specific hyperplasia. Gene expression of peroxisome proliferator-activated receptor γ (PPARG), CCAAT/enhancer-binding protein α (CEBPA), sterol regulatory element-binding transcription factor 1 (SREBF1), wingless-type MMTV integration site family 10B (WNT10B), fatty acid-binding protein 4 (FABP4), acetyl Co-A carboxylase α, lipoprotein lipase and fatty acid synthase (FASN) were determined by real-time quantitative PCR. Expression did not differ between the experimental groups within the tissues but did differ between the tissues: PPARG, FABP4 and FASN were upregulated in the SC AT, while CEBPA, WNT10B and SREBF1 were upregulated in the LM. Although age and diet energy density did not have a significant effect on increasing the amount of IMF, these factors could have influenced adipocyte development in this tissue differently than in the SC AT. This was evidenced by the different size distributions of the cells in the two tissues, and the differing expression patterns of certain markers in the SC AT and the LM, which may indicate a differential role of PPARG and WNT10B in triggering adipocyte proliferation and fat accumulation capacity.

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Section: Methodsmentioning

confidence: 99%

Expression of genes involved in adipogenesis and lipid metabolism in subcutaneous adipose tissue and longissimus muscle in low-marbled Pirenaica beef cattle

Soret

Mendizábal

Arana

et al. 2016

animal

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show abstract

“…Затем крыс усыпляли СО 2 -ас-фиксией, получали сыворотку крови и ýпидиди-мальную жировую ткань, из которой выделяли адипоциты с использованием коллагеназы (Sigma Aldrich, США) по методу M. Rodbell [10]. Жиз-неспособность выделенных клеток оценивали с трипановым синим (Serva, США), при ýтом доля живых клеток составляла не менее 95%.…”

Section: материал и методыunclassified

Oxidative stress in the pathogenesis of type 1 diabetes: the role of adipocyte xanthine oxidase

Иванов¹,

Шахристова²,

Степовая³

et al. 2017

Bûll. sib. med.

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“…These cells were given the name Processed Lipoaspirate or PLA cells due to their derivation from processed lipoaspirate tissue obtained through cosmetic surgery. The isolation method described in this article was based on existing enzymatic strategies for the isolation of the stromal vascular fraction (SVF) from adipose tissue 2 . The SVF has been defined as a minimally processed population of red blood cells, fibroblasts, endothelial cells, smooth muscle cells, pericytes and pre-adipocytes that have yet to adhere to a tissue culture substrate 2,3 .…”

Section: Introductionmentioning

confidence: 99%

“…The isolation method described in this article was based on existing enzymatic strategies for the isolation of the stromal vascular fraction (SVF) from adipose tissue 2 . The SVF has been defined as a minimally processed population of red blood cells, fibroblasts, endothelial cells, smooth muscle cells, pericytes and pre-adipocytes that have yet to adhere to a tissue culture substrate 2,3 . Culturing of this SVF over time is proposed to eliminate many of these contaminating cell populations and result in an adherent, fibroblastic population.…”

Section: Introductionmentioning

confidence: 99%