Leptin and TGF-β1 Downregulate PREP1 Expression in Human Adipose-Derived Mesenchymal Stem Cells and Mature Adipocytes

Adipose tissue is widely recognized as an extremely active endocrine organ producing adipokines as leptin that bridge metabolism and the immune system. Pre-B-cell leukemia homeobox (Pbx)-regulating protein-1 (PREP1) is a ubiquitous homeodomain transcription factor involved in the adipogenic differentiation and insulin-sensitivity processes. Leptin, as pleiotropic adipokine, and TGF-β, known to be expressed by primary pre-adipocytes [adipose-derived stem cells (ASCs)] and mature differentiated adipocytes, modulate inflammatory responses. We aimed to assess for the first time if leptin and TGF-β interfere with PREP1 expression in both ASCs and mature differentiated adipocytes. Human ASCs were isolated from subcutaneous adipose liposuction and, after expansion, fully differentiated to mature adipocytes. In both ASCs and adipocytes, leptin and TGF-β1 significantly decreased the expression of PREP1, alone and following concurrent Toll-like receptor 4 (TLR4) activation. Moreover, in adipocytes, but not in ASCs, leptin increased TLR4 and IL-33 expression, whereas TGF-β1 enhanced TLR4 and IL-6 expression. Taken together, we provide evidence for a direct regulation of PREP1 by leptin and TGF-β1 in ASCs and mature adipocytes. The effects of leptin and TGF-β1 on immune receptors and cytokines, however, are limited to mature adipocytes, suggesting that modulating immune responses depends on the differentiation of ASCs. Further studies are needed to fully understand the regulation of PREP1 expression and its potential for the development of new therapeutic approaches in obesity-related diseases.

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Hybrid Complexes of High and Low Molecular Weight Hyaluronans Highly Enhance HASCs Differentiation: Implication for Facial Bioremodelling

Cellular Physiology and Biochemistry ◽

10.1159/000485414 ◽

2017 ◽

Vol 44 (3) ◽

pp. 1078-1092 ◽

Cited By ~ 27

Author(s):

Antonietta Stellavato ◽

Marcella La Noce ◽

Luisana Corsuto ◽

Anna Virginia Adriana Pirozzi ◽

Mario De Rosa ◽

...

Keyword(s):

Stem Cells ◽

Molecular Weight ◽

Adipogenic Differentiation ◽

Fat Grafting ◽

Tissue Ischemia ◽

Ppar Γ ◽

Human Ascs ◽

Differentiation And Proliferation ◽

Related Proteins ◽

Subdermal Fat

Background/Aims: Adipose-derived Stem Cells (ASCs) are used in Regenerative Medicine, including fat grafting, recovery from local tissue ischemia and scar remodeling. The aim of this study was to evaluate hyaluronan based gel effects on ASCs differentiation and proliferation. Methods: Comparative analyses using high (H) and low (L) molecular weight hyaluronans (HA), hyaluronan hybrid cooperative complexes (HCCs), and high and medium cross-linked hyaluronan based dermal fillers were performed. Human ASCs were characterized by flow cytometry using CD90, CD34, CD105, CD29, CD31, CD45 and CD14 markers. Then, cells were treated for 7, 14 and 21 days with hyaluronans. Adipogenic differentiation was evaluated using Oil red-O staining and expression of leptin, PPAR-γ, LPL and adiponectin using qRT-PCR. Adiponectin was analyzed by immunofluorescence, PPAR-γ and adiponectin were analyzed using western blotting. ELISA assays for adiponectin and leptin were performed. Results: HCCs highly affected ASCs differentiation by up-regulating adipogenic genes and related proteins, that were also secreted in the culture medium. H-HA and L-HA induced a lower level of ASCs differentiation. Conclusion: HCCs-based formulations clearly enhance adipogenic differentiation and proliferation, when compared with linear HA and cross-linked hyaluronans. Injection of HCCs in subdermal fat compartment may recruit and differentiate stem cells in adipocytes, and considerably improving fat tissue renewal.

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282 ADIPOGENIC DIFFERENTIATION IN VITRO OF PORCINE ADULT MESENCHYMAL STEM CELLS

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab282 ◽

2009 ◽

Vol 21 (1) ◽

pp. 238 ◽

Cited By ~ 1

Author(s):

E. Monaco ◽

A. Lima ◽

S. Wilson ◽

S. Lane ◽

M. Bionaz ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Bone Marrow ◽

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Adipogenic Differentiation ◽

Expression Patterns ◽

Mrna Abundance ◽

Subcutaneous Adipose

The quantity and accessibility of subcutaneous adipose tissue in humans make it an attractive alternative to bone marrow as a source of adult stem cells for therapeutic purposes. However, before such a cell source substitution can be proposed, the properties of stem cells derived from adipose tissue (ADSC) and bone marrow (BMSC), and their differentiated progeny must be compared in an animal model, such as swine, that adequately simulates the structure and physiology of humans. The objective of this work was to induce adult porcine stem cells isolated from subcutaneous adipose tissue and bone marrow to differentiate in vitro along the adipogenic lineage and to compare their transcript profile properties. ADSC and BMSC were isolated from subcutaneous adipose tissue and femurs of adult pigs, respectively, and differentiated along the adipogenic lineage using specific inducing medium. Cells were incubated up to 4 weeks with medium replaced every 3 days. Histological staining with Oil Red O was performed at 0, 2, 4, 7, 14, 21, 28 days of differentiation (dd) to confirm the adipogenic differentiation. RNA was also extracted at these time points. qPCR was performed on PPARG, DBI, ACSL1, CD36, CEBPA, DGAT2, ADFP, ADIPOQ, SCD. The geometrical mean of GTF2H3, NUBP, and PPP2CB was used as an internal control. Gene expression was analyzed using a mixed model of SAS with repeated time. The adipogenic differentiation of both ADSC and BMSC was confirmed by the Oil Red O positive staining. The relative mRNA abundance of all the genes at dd0 was similar between the ADSC and BMSC. The relative mRNA abundance of most of the genes was also similar between ADSC and BMSC throughout the adipogenic differentiation. ACSL1 and ADIPOQ had analogous expression patterns among the cell types. ACSL1 had relatively large mRNA abundance before differentiation, but ADIPOQ was barely detectable. As a consequence of differentiation, ACSL1 increased in relative mRNA abundance about 10-fold, whereas ADIPOQ mRNA increased about 1000-fold. Temporal expression patterns of SCD, DGAT2, and ADFP were similar. The increase in gene expression was >800% for SCD, >500% for ADFP, and >50 000% for DGAT2 after 7dd. ADSC had significantly higher expression of those genes compared to BMSC at 14 and 28dd. Both ADIPOQ and DGAT2 were almost undetectable prior to differentiation. mRNA expression of CD36 and DBI was similar with a significantly larger increase in expression of ADSC compared with BMSC. Relative mRNA abundance of CEBPA and PPARG was also larger in ADSC compared with BMSC; however, BMSC had a remarkable increase in temporal expression of those genes throughout adipogenic differentiation. These results suggest both cell types can differentiate towards the adipogenic lineage but with quantitatively different gene expression patterns. More investigation is needed before the ADSC can be considered a practical alternative source for stem cells in future human clinical applications. This research was supported by the Illinois Regenerative Medicine Institute.

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Subcutaneous Adipose Stem Cells in Obesity: The Impact of Bariatric Surgery

10.5772/intechopen.95542 ◽

2021 ◽

Author(s):

Veronica Mocanu ◽

Daniel V. Timofte ◽

Ioana Hristov

Keyword(s):

Insulin Resistance ◽

Bariatric Surgery ◽

Stem Cells ◽

Weight Loss ◽

Adipose Tissue ◽

Adipogenic Differentiation ◽

Low Grade ◽

Potential Weight ◽

Subcutaneous Adipose ◽

The Impact

Adipocyte expansion, which involves adipose tissue-derived mesenchymal stem cells (ASCs), is a critical process with implications in the pathogenesis of metabolic syndrome and insulin resistance associated with obesity. Impaired subcutaneous adipogenesis leads to dysfunctional, hypertrophic adipocytes, chronic low-grade inflammation, and peripheric insulin resistance. Alternatively, it has also been proposed that the preservation of the functionality of subcutaneous adipocyte precursors could contribute to some obese individuals remaining metabolically healthy. Very few studies evaluated the changes in the adipogenic differentiation for human subcutaneous ASCs following bariatric surgery. Weight loss after bariatric surgery involves extensive remodeling of adipose tissue, comprising the hyperplasia-hypertrophy balance. Subcutaneous ASCs may be implicated in the variations of bariatric outcomes, through a different restoration in their proliferative and adipogenic potential. Weight loss induced by bariatric surgery correlates to the subcutaneous ASC functions and could explain the variability of metabolic improvement. Limited research data are available to the present and these data support the importance of diagnosis of subcutaneous ASCs functions as predictors of metabolic improvement after bariatric surgery.

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Epigenetic regulation of Neuregulin-1 tunes white adipose stem cell differentiation

10.1101/2020.03.23.004275 ◽

2020 ◽

Author(s):

Alyssa D. Cordero ◽

Evan C. Callihan ◽

Rana Said ◽

Yasir Alowais ◽

Emily S. Paffhausen ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Adipogenic Differentiation ◽

Stem Cell Differentiation ◽

Metabolic Dysfunction ◽

Human Stem Cells ◽

Adipose Stem Cell ◽

Neuregulin 1 ◽

Subcutaneous Adipose

AbstractExpansion of subcutaneous adipose tissue by differentiation of new adipocytes has been linked to improvements in metabolic health. However, an expandability limit has been observed wherein new adipocytes cannot be produced, the existing adipocytes become enlarged (hypertrophic) and lipids spill over into ectopic sites. Inappropriate ectopic storage of these surplus lipids in liver, muscle, and visceral depots has been linked with metabolic dysfunction. Here we show that Neuregulin-1 (NRG1) serves as a regulator of adipogenic differentiation in subcutaneous primary human stem cells. We further demonstrate that DNA methylation modulates NRG1 expression in these cells, and a 3-day exposure of stem cells to a recombinant NRG1 peptide fragment is sufficient to reprogram adipogenic cellular differentiation to higher levels. These results define a novel molecular adipogenic rheostat with potential implications for the expansion of adipose tissue in vivo.

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Enhanced Adipogenic Differentiation of Human Adipose-Derived Stem Cells in an in vitro Microenvironment: The Preparation of Adipose-Like Microtissues Using a Three-Dimensional Culture

Cell Medicine ◽

10.3727/215517916x693096 ◽

2017 ◽

Vol 9 (1-2) ◽

pp. 35-44 ◽

Cited By ~ 6

Author(s):

Yoshitaka Miyamoto ◽

Masashi Ikeuchi ◽

Hirofumi Noguchi ◽

Tohru Yagi ◽

Shuji Hayashi

Keyword(s):

Stem Cells ◽

Adipogenic Differentiation ◽

Three Dimensional ◽

3D Culture ◽

Human Stem Cells ◽

Maintenance Medium ◽

3D Cultures ◽

Differentiation Medium ◽

Human Ascs

The application of stem cells for cell therapy has been extensively studied in recent years. Among the various types of stem cells, human adipose tissue-derived stem cells (ASCs) can be obtained in large quantities with relatively few passages, and they possess a stable quality. ASCs can differentiate into a number of cell types, such as adipose cells and ectodermal cells. We therefore focused on the in vitro microenvironment required for such differentiation and attempted to induce the differentiation of human stem cells into microtissues using a microelectromechanical system. We first evaluated the adipogenic differentiation of human ASC spheroids in a three-dimensional (3D) culture. We then created the in vitro microenvironment using a 3D combinatorial TASCL device and attempted to induce the adipogenic differentiation of human ASCs. The differentiation of human ASC spheroids cultured in maintenance medium and those cultured in adipocyte differentiation medium was evaluated via Oil red O staining using lipid droplets based on the quantity of accumulated triglycerides. The differentiation was confirmed in both media, but the human ASCs in the 3D cultures contained higher amounts of triglycerides than those in the 2D cultures. In the short culture period, greater adipogenic differentiation was observed in the 3D cultures than in the 2D cultures. The 3D culture using the TASCL device with adipogenic differentiation medium promoted greater differentiation of human ASCs into adipogenic lineages than either a 2D culture or a culture using a maintenance medium. In summary, the TASCL device created a hospitable in vitro microenvironment and may therefore be a useful tool for the induction of differentiation in 3D culture. The resultant human ASC spheroids were “adipose-like microtissues” that formed spherical aggregation perfectly and are expected to be applicable in regenerative medicine as well as cell transplantation.

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Epigenetic Regulation of Neuregulin-1 Tunes White Adipose Stem Cell Differentiation

Cells ◽

10.3390/cells9051148 ◽

2020 ◽

Vol 9 (5) ◽

pp. 1148

Author(s):

Alyssa D. Cordero ◽

Evan C. Callihan ◽

Rana Said ◽

Yasir Alowais ◽

Emily S. Paffhausen ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Adipogenic Differentiation ◽

Stem Cell Differentiation ◽

Metabolic Dysfunction ◽

Human Stem Cells ◽

Adipose Stem Cell ◽

Neuregulin 1 ◽

Subcutaneous Adipose

Expansion of subcutaneous adipose tissue by differentiation of new adipocytes has been linked to improvements in metabolic health. However, an expandability limit has been observed wherein new adipocytes cannot be produced, the existing adipocytes become enlarged (hypertrophic) and lipids spill over into ectopic sites. Inappropriate ectopic storage of these surplus lipids in liver, muscle, and visceral depots has been linked with metabolic dysfunction. Here we show that Neuregulin-1 (NRG1) serves as a regulator of adipogenic differentiation in subcutaneous primary human stem cells. We further demonstrate that DNA methylation modulates NRG1 expression in these cells, and a 3-day exposure of stem cells to a recombinant NRG1 peptide fragment is sufficient to reprogram adipogenic cellular differentiation to higher levels. These results define a novel molecular adipogenic rheostat with potential implications for the expansion of adipose tissue in vivo.

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P0-Related Protein Accelerates the Migration of Human Mesenchymal Stem Cells and Their Progeny by Modulating VLA-5 Interactions with Fibronectin.

Blood ◽

10.1182/blood.v108.11.1391.1391 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1391-1391

Author(s):

Suzanne M. Watt ◽

Maria Roubelakis ◽

Sinead Forde ◽

Fengjuan Lu ◽

Grigorios Tsaknakis ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Inflammatory Responses ◽

Human Mesenchymal Stem Cells ◽

Leading Edge ◽

Cancer Surveillance ◽

Related Protein ◽

Transmembrane Receptors ◽

Type Iv ◽

First Time

Abstract Human mesenchymal stem cells (hMSCs) are capable of expansion, self renewal and generation of several tissues such as bone, fat and cartilage. They also generate microenviromental niche cells that regulate angiogenesis and hematopoiesis. During development and in the adult, hMSCs and their progeny migrate to specific niches where they provide support for hematopoietic cells or they play an important role in tissue regeneration, inflammatory responses and cancer surveillance. There is particular interest in understanding the molecular basis of hMSCs migration which involves numerous transmembrane receptors, cell adhesion and signaling molecules. Here, we demonstrate, for the first time, that human P0 - related protein, hPZR, has the ability to accelerate VLA-5-mediated migration of cultured hMSCs on the extracellular matrix (ECM) protein, fibronectin. This is mediated via the immunotyrosine-inhibitory motif (ITIM) sequences within the hPZR cytoplasmic domain, and the ECM ligand, since it cannot be recapitulated with the hPZR ITIM-less isoform, hPZRb, nor with other ECM components such as laminin or Type IV collagen. It is mediated by interactions with the docking phosphatase, SHP-2. The specificity of these results was confirmed by knocking-down hPZR and hPZRb using siRNA technology. We further demonstrated that hPZR clusters with VLA-5 at the leading edge of the cell, using high-resolution confocal microscopy together with immunoprecipitation and immunoblotting technologies. We propose that hPZR plays a key role in negatively regulating VLA-5 adhesion to fibronectin, a process critical for cell detachment from fibronectin and for the migration of hMSCs and their progeny.

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GABA-stimulated adipose-derived stem cells suppress subcutaneous adipose inflammation in obesity

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1822067116 ◽

2019 ◽

pp. 201822067 ◽

Cited By ~ 1

Author(s):

Injae Hwang ◽

Kyuri Jo ◽

Kyung Cheul Shin ◽

Jong In Kim ◽

Yul Ji ◽

...

Keyword(s):

Insulin Resistance ◽

Stem Cells ◽

Adipose Tissue ◽

Inflammatory Responses ◽

Epididymal Adipose Tissue ◽

Adipose Derived Stem Cells ◽

Tissue Macrophage ◽

Monocyte Migration ◽

Fat Depot ◽

Subcutaneous Adipose

Accumulating evidence suggests that subcutaneous and visceral adipose tissues are differentially associated with metabolic disorders. In obesity, subcutaneous adipose tissue is beneficial for metabolic homeostasis because of repressed inflammation. However, the underlying mechanism remains unclear. Here, we demonstrate that γ-aminobutyric acid (GABA) sensitivity is crucial in determining fat depot-selective adipose tissue macrophage (ATM) infiltration in obesity. In diet-induced obesity, GABA reduced monocyte migration in subcutaneous inguinal adipose tissue (IAT), but not in visceral epididymal adipose tissue (EAT). Pharmacological modulation of the GABAB receptor affected the levels of ATM infiltration and adipose tissue inflammation in IAT, but not in EAT, and GABA administration ameliorated systemic insulin resistance and enhanced insulin-dependent glucose uptake in IAT, accompanied by lower inflammatory responses. Intriguingly, compared with adipose-derived stem cells (ADSCs) from EAT, IAT-ADSCs played key roles in mediating GABA responses that repressed ATM infiltration in high-fat diet-fed mice. These data suggest that selective GABA responses in IAT contribute to fat depot-selective suppression of inflammatory responses and protection from insulin resistance in obesity.

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Tissue Source and Cell Expansion Condition Influence Phenotypic Changes of Adipose-Derived Stem Cells

Stem Cells International ◽

10.1155/2017/7108458 ◽

2017 ◽

Vol 2017 ◽

pp. 1-15 ◽

Cited By ~ 7

Author(s):

Lauren H. Mangum ◽

Shanmugasundaram Natesan ◽

Randolph Stone ◽

Nicole L. Wrice ◽

David A. Larson ◽

...

Keyword(s):

Stem Cells ◽

Culture Conditions ◽

Adipose Derived Stem Cells ◽

Burned Skin ◽

Human Ascs ◽

Triglyceride Content ◽

Human Platelet Lysate ◽

Surface Staining ◽

Expansion Condition ◽

Subcutaneous Adipose

Stem cells derived from the subcutaneous adipose tissue of debrided burned skin represent an appealing source of adipose-derived stem cells (ASCs) for regenerative medicine. Traditional tissue culture uses fetal bovine serum (FBS), which complicates utilization of ASCs in human medicine. Human platelet lysate (hPL) is one potential xeno-free, alternative supplement for use in ASC culture. In this study, adipogenic and osteogenic differentiation in media supplemented with 10% FBS or 10% hPL was compared in human ASCs derived from abdominoplasty (HAP) or from adipose associated with debrided burned skin (BH). Most (95–99%) cells cultured in FBS were stained positive for CD73, CD90, CD105, and CD142. FBS supplementation was associated with increased triglyceride content and expression of adipogenic genes. Culture in hPL significantly decreased surface staining of CD105 by 31% and 48% and CD142 by 27% and 35% in HAP and BH, respectively (p<0.05). Culture of BH-ASCs in hPL also increased expression of markers of osteogenesis and increased ALP activity. These data indicate that application of ASCs for wound healing may be influenced by ASC source as well as culture conditions used to expand them. As such, these factors must be taken into consideration before ASCs are used for regenerative purposes.

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