Cell Models and Their Application for Studying Adipogenic Differentiation in Relation to Obesity: A Review

Ruiz‐Ojeda, Francisco Javier; Rupérez, Azahara I.; Gómez-Llorente, Carolina; Gil, Ángel; Aguilera, Concepción M.

doi:10.3390/ijms17071040

Cited by 273 publications

(222 citation statements)

References 170 publications

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“…Furthermore, chemicals or small molecules including cytokines are major factors in the induction of cell differentiation [25][26][27][28]. Francisco et al found that human preadipocytes and ASCs were excellent models for studying adipogenesis and obesity-related metabolic alterations and were appropriate for revealing adipocyte renewal, development and depot-specific differences [4]. Therefore, we focused on uncovering the molecular regulation mechanism as to how miRNAs regulate adipogenic differentiation of hASCs.…”

Section: Discussionmentioning

confidence: 99%

“…Monitoring the alteration of adipose-derived stem cells (ASCs) can provide a promising future in the field of tissue engineering and regenerative medicine [3]. First, ASCs are recruited by certain stimulation, following which they are transformed into preadipocytes and then ultimately turn into mature adipocytes [4]. During adipogenesis, several transcription factors such as peroxisome proliferator-activated receptor-γ (PPARγ) and members of the CCAAT/enhancer-binding family of proteins (c/EBPα) participate in regulating adipogenesis [5,6].…”

Section: Introductionmentioning

confidence: 99%

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MiR-27b Impairs Adipocyte Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells by Targeting LPL

Hu¹,

Tang²,

Hu³

et al. 2018

Cell Physiol Biochem

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Background/Aims: In this study, the molecular mechanisms of miR-27b and lipoprotein lipase (LPL) that regulate human adipose-derived mesenchymal stem cells (hASCs) adipogenic differentiation were detected. Methods: Microarray analysis was applied to screen for differentially expressed miRNAs and mRNA during hASCs adipocyte differentiation induction. MiR-27b and LPL were found to have abnormal expression. Then, a dual luciferase reporter assay was employed to validate the targeting relationship between miR-27b and LPL. We also utilized qRT-PCR, western blot, cellular immunofluorescence and an oil red O staining assay to analyze the regulation of miR-27b and LPL during adipogenic differentiation. Results: The microarray analysis demonstrated that, during adipogenic differentiation, miR-27b was down-regulated, while LPL was up-regulated but tended to become stable 14 days after induction. A dual luciferase reporter assay confirmed the negative targeting regulatory relationship between miR-27b and LPL. After overexpressing and silencing miR-27b, LPL was found to be reversely regulated by miR-27b according to qRT-PCR and western blot. The fat-formation-related biomarkers CCAAT-enhancer binding protein α (c/EBPα) and peroxisome proliferator-activated receptors γ (PPARγ) had decreasing levels after over-expressing miR-27b or knockdown of LPL followed by adipogenic differentiation. Meanwhile, the oil red O staining assay revealed that the accumulation of lipid droplets decreased. There was no change in the expression of c/EBPα, PPARγ, or lipid droplet accumulation when overexpressing miR-27b and LPL. Conclusion: During the adipogenic differentiation of hASCs, miR-27b expression decreased, and LPL expression increased. The abnormal expression of miR-27b and LPL effectively regulated the adipogenic differentiation of hASCs.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

MiR-27b Impairs Adipocyte Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells by Targeting LPL

Hu¹,

Tang²,

Hu³

et al. 2018

Cell Physiol Biochem

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“…The 3T3-L1 is a cell line derived from mouse 3T3 cells, which is an adipocyte with fibroblast like morphology (Ruiz-Ojeda, Ruperez, Gomez-Llorente, Gil, & Aguilera, 2016). The fibroblast cell lines viz.…”

Section: In Vitro Wound Healing Assaymentioning

confidence: 99%

Untitled

Kumar¹,

Shakila²,

Jeyasekaran³

2019

Turk. J. Fish. Aquat. Sci.

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In this study, skin of unicorn leatherjacket (Aluterus monoceros) was hydrolyzed using collagenase at three different temperatures viz. 5°C, 25°C and 50°C, to obtain collagen hydrolysates. Collagen peptides of low molecular masses (<3kDa) viz. CP-5, CP-25 and CP-50 were obtained by sequential ultrafiltration with tangential flow filtration (TFF) system. The results indicated that CP-5 exhibited the maximum in vitro anti-cancer, anti-diabetic and wound healing activities than CP-25 and CP-50. In MTT assay, CP-5 inhibited the viability of COLO320 cancer cells upto 49.78% at 1 mg/mL concentration, while the inhibition was only 29.92% by CP-50. Anti-diabetic activity as assessed by α-amylase inhibition was 80.45% by CP-5 at 1 mg/mL concentration, as against 75.81% and 71.17% by CP-25 and CP-50, respectively. Collagen peptides, in general, exhibited very low anti-inflammatory property, which ranged between 3.27% to 4.01%. In vitro wound healing tested by scratch assay showed good migration of 3T3-L1 cells in CP-5 treated than CP-25 and CP-50 at 0.2 mg/mL concentration. The study thus proved that collagen peptides derived from fish skin at low hydrolysis temperatures (50C) exhibited more bio-active activities than those hydrolysed at high temperatures (500C); and shall be effectively utilized in various bio-medical applications.

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“…Results from these studies are not as useful for applications on human health as expected, given the physiological and metabolic differences among species [2]. Human primary cultured cells are the best resource to understand human adipocyte biology in deranged metabolic states [2], but cannot inform about cell events that occur during prenatal life.…”

Section: Introductionmentioning

confidence: 99%

PPARγ agonist through the terminal differentiation phase is essential for adipogenic differentiation of fetal ovine preadipocytes

Pu¹,

Veiga-López²

2017

Cell Mol Biol Lett

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BackgroundAlthough the 3T3-L1 preadipocyte cell line represents an informative model for in vitro adipogenesis research, primary cultured cells are often needed to understand particular human or animal metabolic phenotypes. As demonstrated by in vitro cultured preadipocytes from large mammalian species, primary cultured cells require specific adipogenic differentiation conditions different to that of the 3T3-L1 cell line. These conditions are also species-specific and require optimization steps. However, efficient protocols to differentiate primary preadipocytes using alternative species to rodents are scarce. Sheep represent an amenable animal model for fetal biology and developmental origins of health and disease studies. In this work, we present with the first detailed procedure to efficiently differentiate primary fetal and adult ovine preadipocytes.MethodsFetal and adult ovine adipose and skin tissue harvest, preadipocyte and fibroblast isolation, proliferation, and standardization and optimization of a new adipogenic differentiation protocol. Use of commercial cell lines (3T3-L1 and NIH-3T3) for validation purposes. Oil red O stain and gene expression were used to validate adipogenic differentiation. ANOVA and Fisher’s exact test were used to determine statistical significance.ResultsOur optimized adipogenic differentiation method included a prolonged adipogenic cocktail exposure time from 2 to 8 days, higher insulin concentration, and supplementation with the peroxisome proliferator-activated receptor gamma (PPARγ) agonist, rosiglitazone. This protocol was optimized for both, fetal and adult preadipocytes.ConclusionsOur protocol enables successful adipogenic differentiation of fetal and adult ovine preadipocytes. This work demonstrates that compared to the 3T3-L1 cell line, fetal ovine preadipocytes require a longer exposure to the differentiation cocktail, and the need for IMBX, dexamethasone, and/or the PPARγ agonist rosiglitazone through the terminal differentiation phase. They also require higher insulin concentration during differentiation to enhance lipid accumulation and similar to human primary preadipocytes, PPARγ agonist supplementation is also required for ovine adipogenic differentiation. This work highlights species-specific differences requirements for adipogenic differentiation and the need to develop standardized methods to investigate comparative adipocyte biology.

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Cell Models and Their Application for Studying Adipogenic Differentiation in Relation to Obesity: A Review

Cited by 273 publications

References 170 publications

MiR-27b Impairs Adipocyte Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells by Targeting LPL

MiR-27b Impairs Adipocyte Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells by Targeting LPL

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PPARγ agonist through the terminal differentiation phase is essential for adipogenic differentiation of fetal ovine preadipocytes

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