The influence of extracellular matrix substrata on preadipocyte development in serum-free cultures of stromal-vascular cells.

Hausman, G J; Wright, J.R.; Richardson, R. L.

doi:10.2527/1996.7492117x

Cited by 59 publications

(53 citation statements)

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“…Our study is the first to use gelatin for pre-adipocyte attachment and proliferation in fish. Most studies in mammals have used laminin as substrate in primary cell culture, as in porcine stromal-vascular cultures, in which laminin increases the number, size, and proportion of adipocytes (Hausman et al 1996). Other studies have described that differentiating pre-adipocytes show greater adherence to laminin than nonpre-adipocytes.…”

Section: Discussionmentioning

confidence: 99%

Regulation of proliferation and differentiation of adipocyte precursor cells in rainbow trout (Oncorhynchus mykiss)

Bouraoui¹,

Gutiérrez²,

Navarro³

2008

Journal of Endocrinology

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Here, we describe optimal conditions for the culture of rainbow trout (Oncorhynchus mykiss) pre-adipocytes obtained from adipose tissue and their differentiation into mature adipocytes, in order to study the endocrine control of adipogenesis. Pre-adipocytes were isolated by collagenase digestion and cultured on laminin or 1% gelatin substrate. The expression of proliferating cell nuclear antigen was used as a marker of cell proliferation on various days of culture. Insulin growth factor-I stimulated cell proliferation especially on days 5 and 7 of culture. Tumor necrosis factor a (TNFa) slightly enhanced cell proliferation only at a low dose. We verified the differentiation of cells grown in specific medium into mature adipocytes by oil red O (ORO) staining. Quantification of ORO showed an increase in triglycerides throughout culture.Immunofluorescence staining of cells at day 11 revealed the expression of CCAAT/enhancer-binding protein and peroxisome proliferator-activator receptor g, suggesting that these transcriptional factors are involved in adipocyte differentiation in trout. We also examined the effect of TNFa on the differentiation of these adipocytes in primary culture. TNFa inhibited the differentiation of these cells, as indicated by a decrease in glycerol-3-phosphate dehydrogenase activity, an established marker of adipocyte differentiation. In conclusion, the culture system described here for trout pre-adipocytes is a powerful tool to study the endocrine regulation of adipogenesis in this species.

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Section: Discussionmentioning

confidence: 99%

Regulation of proliferation and differentiation of adipocyte precursor cells in rainbow trout (Oncorhynchus mykiss)

Bouraoui¹,

Gutiérrez²,

Navarro³

2008

Journal of Endocrinology

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“…This hormone has also been shown to regulate stearoyl-CoA the synthesis of ECM components. Indeed, dexamethasone was reported to enhance the production of laminin and type IV collagen in porcine primary preadipocytes [113,115]. These ECM components are critical for the development of the adipocytes that are embedded in a basement membrane.…”

Section: Ligands For Nuclear Receptorsmentioning

confidence: 99%

The adipose conversion process: regulation by extracellular and intracellular factors

Boone

Mourot

Gregoire³

et al. 2000

Reprod. Nutr. Dev.

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-White adipose tissue regulates lipid metabolism and acts as a secretory organ. Because of its importance for human health and animal production, many studies have attempted to better understand its development at the cellular and molecular levels by culturing preadipose cells in vitro. This synthesis article describes our current knowledge, acquired by this approach, concerning the regulation of the different steps of the adipocyte differentiation program by extracellular (hormones, cytokines, growth factors, retinoids and fatty acids) and intracellular agents (second messengers and transcription factors). The discrepant effects that have been observed for some of these factors are also discussed. This information is very important in the perspective of a better control of fat deposits in human and breeding species.preadipocyte / differentiation / hormonal agent / second messenger / transcription factor Résumé -Le processus d'adipoconversion : sa régulation par des facteurs extracellulaires et intracellulaires. Le tissu adipeux blanc régule le métabolisme lipidique et agit comme un organe de sécrétion. Étant donné son importance pour la santé humaine et la production animale, de nombreuses études ont tenté de mieux comprendre son développement aux niveaux cellulaire et moléculaire en utilisant des cultures de préadipocytes. Cet article de synthèse décrit notre connaissance actuelle, issue de cette approche, concernant la régulation des différentes étapes du programme de la diffé-renciation adipocytaire par des facteurs extracellulaires (hormones, cytokines, facteurs de croissance, rétinoïdes et acides gras) et intracellulaires (seconds messagers et facteurs de transcription). Les effets divergents observés pour certains de ces facteurs sont également discutés. Ces informations sont très importantes dans la perspective d'un meilleur contrôle des dépôts adipeux chez l'humain et les espèces d'élevage.préadipocyte / différenciation / hormone / second messager / facteur de transcription Reprod. Nutr. Dev. 40 (2000) 325-358 325

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“…Cell adhesion has been strongly implicated in the regulation of adipogenesis. In addition to the cell rounding in the first 24 h of adipogenesis (Spiegelman and Farmer, 1982;Hausman et al, 1996), enhanced adhesion of 3T3-L1 cells on fibronectin diminishes differentiation (Castro-Munozledo et al, 1987;Kamiya et al, 2002). The actinbinding protein enc1 is involved in both morphology changes and differentiation during adipogenesis (Zhao et al, 2000).…”

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confidence: 99%

2,3,7,8-Tetrachlorodibenzo-p-dioxin and Epidermal Growth Factor Cooperatively Suppress Peroxisome Proliferator-Activated Receptor-γ1 Stimulation and Restore Focal Adhesion Complexes during Adipogenesis: Selective Contributions of Src, Rho, and Erk Distinguish These Overlapping Processes in C3H10T1/2 Cells

Liu

Jefcoate²

2006

Mol Pharmacol

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Stimulation of PPAR␥1 and adipogenesis in multipotential C3H10T1/2 cells by the combination of dexamethasone and 3-isobutyl-1-methylxanthine (DM) is suppressed by 2,3,7,8 tetrachlorodibenzodioxin (TCDD) (10 nM). This suppression requires sustained activation of extracellular signal-regulated kinase (Erk)1/2. We show that it arises from an effect of TCDD on epidermal growth factor (EGF) signaling. DM initiates an early loss of cell adhesion that is reversed by this TCDD/EGF synergy. Src kinase activity was completely essential for adhesion restoration, sustained Erk activation, and suppression of peroxisome proliferator-activated receptor (PPAR)␥1. MEK/Erk activity did not contribute, however, to TCDD-induced adhesion. Stimulation of adhesion may therefore precede elevation of Erk. Adhesion is produced by interaction of ␣␤ integrins with extracellular matrix proteins and subsequent Src-mediated phosphorylation of focal adhesion kinase (FAK, Tyr576/577) and paxillin (Tyr118). TCDD enhanced the steady state Src-mediated phosphorylation of FAK but not of paxillin. Protein tyrosine phosphatase (PTPase) inhibition by orthovanadate (OVA) showed that this Src activity is highly restricted by PTPases. Partial inhibition of PTPases by OVA mimicked TCDD in producing EGF-and Src-dependent effects on cell adhesion and PPAR␥1 suppression. TCDD may therefore induce a protein that enhances Src effectiveness at adhesion sites. Rho kinase (ROCK) inhibition blocked TCDD/EGF stimulation of clustered focal adhesion complexes without affecting either sustained Erk activation or suppression of PPAR␥1. Thus, this ROCKmediated clustering of integrin complexes is not needed for the effects of TCDD on Erk and PPAR␥1. A minimal cholesterol depletion with ␤-methylcyclodextrin attenuated TCDD effects on PPAR␥1 and Erk activation. TCDD intervention is therefore linked to extracellular proteins. It indicates that TCDD-enhanced stimulation of EGF signaling to Erk may derive from the initial ␣␤ integrin complexes.Extensive studies on adipocyte differentiation have used mouse embryonic fibroblastic cell models, including 3T3-L1 and 3T3-422A, which are already committed to the adipocyte lineage. C3H10T1/2 (10T1/2), a pluripotent progenitor cell type that is similar to mouse embryo fibroblasts, has also been used (Alexander et al., 1998). This cell type develops into fat, bone, or muscle cells according to the stimulus (Ntambi and Young-Cheul, 2000). In general, differentiation of 3T3-L1 and 10T1/2 cells is stimulated by a hormonal mixture (IDM) composed of insulin, dexamethasone (Dex), and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), which is administered to confluent cells along with serum renewal (Cowherd et al., 1999). Over approximately 48 h in 10T1/2 cells, IDM cooperatively directs several Article, publication date, and citation information can be found at

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The influence of extracellular matrix substrata on preadipocyte development in serum-free cultures of stromal-vascular cells.

Cited by 59 publications

References 0 publications

Regulation of proliferation and differentiation of adipocyte precursor cells in rainbow trout (Oncorhynchus mykiss)

Regulation of proliferation and differentiation of adipocyte precursor cells in rainbow trout (Oncorhynchus mykiss)

The adipose conversion process: regulation by extracellular and intracellular factors

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