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Adipogenesis

Adipogenesis is the process of cell differentiation by which preadipocytes become mature adipocytes.
This process involves the accumulation of lipid droplets, changes in gene expression and morphology, and the acquisition of the characteristic features of adipocytes.
Adipogenesis is an important aspect of adipose tissue development and is regulated by a complex network of transcription factors, signaling pathways, and epigenetic modifications.
Understanding the mechanisms of adipogenesis is crucial for studying obesity, diabetes, and other metabolic disorders.
Researchers utilize various in vitro and in vivo models to investigate the molecular and cellular events underlying adipocyte differentiation and function.

Most cited protocols related to «Adipogenesis»

Adipogenesis Transcriptional Regulation Analysis

For differentially expressed genes from RNA-seq data, gene ontology (GO) analysis was carried out using DAVID (https://david.ncifcrf.gov). For ChIP-seq data analysis with spike-in normalization, sequences were aligned to either mouse genome (mm9) or Drosophila genome (dm6). Normalization factors were determined by counting Drosophila tags and applying to mouse tags to generate normalized heat maps and profiles. To identify H3K4me1 or H3K27ac enriched regions at day 4 (D4) of adipogenesis in LSL-K4M;CreER brown preadipocytes, we used ‘SICER’ method (25 (link)) with window size of 200 bp and with an estimated false discovery rate (FDR) threshold of 10⁻¹⁰. Previously published ChIP-seq data sets for MLL4, PPARγ, C/EBPα, CBP, BRD4 and MED1 at day 2 (D2) of adipogenesis were used (GSE50466, GSE74189, and GSE99101) (4 (link),6 (link),7 (link)) and the window size was chosen to be 50 bp. To define MLL4⁺ active enhancers during adipogenesis, we compared 13 871 MLL4 binding sites at D2 with 38 618 active enhancers (H3K4me1⁺H3K27ac⁺ at D4). 8777 MLL4⁺ (22.7%, 8777/38 618) sites were located on active enhancers. Heat maps were generated with 50 bp resolution and ranked according to the intensity of 8777 MLL4⁺ active enhancers at the center. Average profiles were plotted using the number of ChIP-seq reads from the center of 8777 MLL4⁺ active enhancers to 5 kb on both sides.

Jang Y., Broun A., Wang C., Park Y.K., Zhuang L., Lee J.E., Froimchuk E., Liu C, & Ge K. (2018). H3.3K4M destabilizes enhancer H3K4 methyltransferases MLL3/MLL4 and impairs adipose tissue development. Nucleic Acids Research, 47(2), 607-620.

Publication 2018

Adipogenesis Binding Sites BRD4 protein, human Chromatin Immunoprecipitation Sequencing Drosophila Genes Genome KMT2B protein, human MED1 protein, human Microtubule-Associated Proteins Mus PPAR gamma RNA-Seq

Optimizing Mouse BM-MSC Isolation Protocols

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Publication 2014

Adipogenesis CD44 protein, human Cell Culture Techniques Cells Chondrogenesis Cultured Cells Endothelial Cells Females Femur Flow Cytometry Hematopoietic System isolation Males Mesenchymal Stem Cells Mus Osteogenesis solvent red 27 Strains Thy-1 Antigens Tolonium Chloride

Assessing Adipose Tissue Transcriptome Variability

To assess the transcript variability in a wide panel of samples we put together a set of 180 samples selected to encompass a broad range of adipose tissue origins and experimental conditions (Table 1). Human fat depots were represented by omental, abdominal subcutaneous, and gluteal tissue. The effect of obesity was considered: lean (BMI <25 kg/m²) vs. obese (BMI >30 kg/m²), with equal gender representation. Growth pattern and stimulation of adipogenesis was represented by including surgically removed lipomas vs. normal adjacent adipose tissue and samples taken before and after 14 days of systemic rosiglitazone treatment (4 mg BD) (11 (link)). Methodological issues like biopsy retrieval method (needle vs. surgical) and RNA extraction method (Tri-reagent vs. column) were also included. Finally we prepared differentiated adipocytes from preadipocytes isolated from the stromovascular fraction of subcutaneous biopsies (Table 1). Needle biopsy samples were taken under local anesthesia using a 12-gauge needle and immediately frozen in liquid nitrogen. Surgical biopsies were taken during elective surgery and immediately frozen. Preadipocytes were differentiated and exposed to either 0 μm, 50 μm, or 200 μm palmitate (13 (link)). All biopsies and cells were homogenized in Tri-reagent (cat. no. AM9738, Ambion, Austin, TX) and RNA was extracted with either a standard Tri-reagent protocol or using Ambion MirVana columns (cat. no. AM1561, Ambion).

Neville M.J., Collins J.M., Gloyn A.L., McCarthy M.I, & Karpe F. (2010). Comprehensive Human Adipose Tissue mRNA and MicroRNA Endogenous Control Selection for Quantitative Real-Time-PCR Normalization. Obesity (Silver Spring, Md.), 19(4), 888-892.

Publication 2010

Abdomen Adipocytes Adipogenesis austin Biopsy Buttocks Cells Elective Surgical Procedures Freezing Genes, vif Homo sapiens Lipoma Local Anesthesia Needle Biopsies Needles Nitrogen Obesity Omentum Operative Surgical Procedures Palmitate Rosiglitazone Tissue, Adipose Tissues

Adipocyte Differentiation Assays Protocols

For adipocyte differentiation assays, confluent cultures of 3T3-L1 and Swiss 3T3 subclones were exposed to induction medium containing dexamethasone (1 μM), insulin (5 μg/ml), and isobutylmethylxanthine (0.5 mM) (DMI) and 10% FBS. 48 hours after induction, cells were maintained in DMEM containing insulin (5 μg/ml) and 10% FBS until ready for harvest. For NIH 3T3 cells, differentiation medium contained DMI, 6% FBS, and 1 μM rosiglitazone. After induction, cells were maintained in medium containing 6% FBS, insulin, and 1 μM rosiglitazone until ready for harvest. For BMP-induced adipogenesis of NIH 3T3 cell lines, cells were grown to confluence, and maintained at post-confluence, in medium containing 6 ng/mL BMP4 or 25 ng/mL BMP2 along with insulin and rosiglitazone. The derivation and genotyping of Zfp423 knockout mice has been previously described 14 (link), 23 (link). All animal experiments were performed according to procedures approved by the Dana-Farber Cancer Institute’s and Beth Isreal Deconess Medical Center’s Institutional Animal Care and Use Committee.

Gupta R.K., Arany Z., Seale P., Mepani R.J., Ye L., Conroe H.M., Roby Y.A., Kulaga H., Reed R.R, & Spiegelman B.M. (2010). Transcriptional Control of Preadipocyte Determination by Zfp423. Nature, 464(7288), 619-623.

Publication 2010

3T3-L1 Cells Adipocytes Adipogenesis Biological Assay BMP2 protein, human Bone Morphogenetic Protein 4 Cells Dexamethasone Institutional Animal Care and Use Committees Insulin Malignant Neoplasms Mice, Knockout NIH 3T3 Cells Rosiglitazone

Adipocyte Differentiation and Thermogenic Activation

Immortalized progenitor cells were plated and grown in DMEM/H medium supplemented with 10% FBS (referred as day 0). For adipocyte differentiation, cell were grown for 6 days until reaching confluence (day 6), and then treated with the adipogenic induction medium as described above for 12 days (day 18). To further stimulate thermogenic program, fully differentiated cells were incubated with 10μM forskolin or 1 μM norepinephrine for 4 h. For BMPs and FGF21 pre-treatment, recombinant BMP7 (3.3 nM), BMP8 (3.3 nM), or FGF21 (50 nM) were added to undifferentiated cells in medium containing insulin (0.5 μM), T3 (2 nM) and 2% FBS for 6 days followed by adipogenic induction for 12 days. For BMPs and FGF21 post-treatment, fully differentiated adipocytes at day 18 were treated with recombinant BMP7 (3.3 nM), BMP8 (3.3 nM), or FGF21 (50 nM) in medium containing insulin (0.5 μM), T3 (2 nM) and 2% FBS for 2 days. We routinely check for mycoplasma contamination and all the cells used in this study are free of mycoplasma.

Xue R., Lynes M.D., Dreyfuss J.M., Shamsi F., Schulz T.J., Zhang H., Huang T.L., Townsend K.L., Li Y., Takahashi H., Weiner L.S., White A.P., Lynes M.S., Rubin L.L., Goodyear L.J., Cypess A.M, & Tseng Y.H. (2015). Clonal analyses and gene profiling identify genetic biomarkers of human brown and white preadipocyte thermogenic potential. Nature medicine, 21(7), 760-768.

Publication 2015

Adipocytes Adipogenesis Bone Morphogenetic Protein 7 Bone Morphogenetic Proteins Cells Colforsin fibroblast growth factor 21 Insulin Mycoplasma Norepinephrine Pancreatic beta Cells Stem Cells Thermogenesis

Most recents protocols related to «Adipogenesis»

Obesity and Adipocyte ACAT Expression

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Example 1

To examine the function of ACATs in obesity, the expression patterns of ACAT1 and ACAT2 genes, and their gene products during adipogenesis of murine 3T3-L1 preadipocytes in vitro were examined. ACAT1 mRNA level was markedly increased in adipocytes from 2 days after initiation of adipogenesis (i.e., D2) as judged by real-time PCR assay (FIG. 1). However, ACAT2 mRNA level was similar between preadipocytes (D0) and mature adipocytes (D6) while a temporal reduction of ACAT2 level was observed at D2 (FIG. 1). In addition, white adipose tissue (WAT) isolated from high fat diet-induced obese mice displayed elevated mRNA level of ACAT1 and reduced mRNA level of ACAT2 when compared with those in lean mice as judged by real-time PCR assay. Leptin level was measured in WAT from lean and obese mice to ensure the development of obesity (FIG. 2). In addition, brown adipose tissue (BAT) from obese mice exhibited elevated levels of both ACAT1 and ACAT2. Uncoupling protein-1 (UCP-1) level was measured in BAT from lean and obese mice as a BAT-specific marker protein (FIG. 2). However, liver from lean and obese mice exhibited similar levels of ACAT1 and ACAT2 (FIG. 2).

US11872198B2. Compositions and methods for regulating body weight and metabolic syndromes (2024-01-16). Purdue Research Foundation [US]. Inventors: Kee-Hong Kim [US], Yuyan Zhu [US].

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Patent 2024

3T3-L1 Cells Adipocytes Adipogenesis a protein, mouse Biological Assay Brown Adipose Tissue Uncoupling Protein Brown Fat CES1 protein, human Diet, High-Fat Genes Leptin Liver Mice, Obese Mus Obesity Proteins Real-Time Polymerase Chain Reaction RNA, Messenger White Adipose Tissue

Avasimibe Inhibits Adipocyte Cholesterol Accumulation

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Example 4

Previous studies reported an adipogenesis-dependent increase in cholesterol accumulation in adipocytes. Similarly, adipogenesis was associated with elevated level free cholesterol with little change in CE level. Moreover, avasimibe treatment resulted in a significantly reduced level of cholesterol and a complete inhibition of CE accumulation in adipocytes during adipogenesis (FIG. 9). This was accompanied by a marked decrease in mRNA levels of genes involved in cholesterol uptake such as scavenger receptor class B member 1 (SR-BI) and CD-36 in adipocytes treated with avasimibe when compared with those in control adipocytes (FIG. 10). Using a 25-[N-[(7-nitro-2-1,3-benzoxadiazol-4-yl)methyl]amino]-27-norcholesterol (25-NBD cholesterol)-based ACAT assay, avasimibe indeed inhibited the ACAT activity, thereby lowering accumulation of fluorescence-labeled CE in adipocytes (FIG. 11). Collectively, the results indicate that avasimibe not only inhibits synthesis of fatty acids and TGs, but also inhibits cholesterol uptake and accumulation of free cholesterol and CE in adipocytes, possibly through suppression of expression of genes involved in cholesterol uptake from the medium.

US11872198B2. Compositions and methods for regulating body weight and metabolic syndromes (2024-01-16). Purdue Research Foundation [US]. Inventors: Kee-Hong Kim [US], Yuyan Zhu [US].

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Patent 2024

25-(N-((7-nitrobenz-2-oxa-1,3-diazol-4-yl)methyl)amino)-27-norcholesterol Adipocytes Adipogenesis Anabolism avasimibe Biological Assay Cardiac Arrest CES1 protein, human Cholesterol Fatty Acids Fluorescence Gene Expression Genes Hypercholesterolemia Metabolism Psychological Inhibition RNA, Messenger Scavenger Receptor

Adipogenic Differentiation of hPDLSCs

Adipogenic differentiation of hPDLSCs cells was determined following the steps of Oil Red O staining kit (Beyotime). The cells were fixed in 4% paraformaldehyde, washed three times with PBS, and covered with an appropriate amount of staining wash buffer for 20 s. Later, the wash buffer was removed and a proper amount of Oil Red O staining solution was added to carry out cell staining for 10–20 min. Afterwards, Oil Red O staining solution was removed, and the right amount of staining wash buffer was added to stand for 30 s. And then, the buffer was removed and the cells were washed with PBS for 20 s. Lastly, an appropriate amount of PBS was added to evenly cover the cells, and the cells were placed under a microscope for observation and photography.

Jiang J., Zhang N., Song H., Yang Y., Li J, & Hu X. (2023). Oridonin alleviates the inhibitory effect of lipopolysaccharide on the proliferation and osteogenic potential of periodontal ligament stem cells by inhibiting endoplasmic reticulum stress and NF-κB/NLRP3 inflammasome signaling. BMC Oral Health, 23, 137.

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Publication 2023

Adipogenesis Buffers Cells Differentiations, Cell Microscopy paraform

Osteogenic and Adipogenic Differentiation of hPDLSCs

Osteogenic differentiation induction was performed on hPDLSCs. When the cell confluence reached about 70%, mineralization induction solution (α-MEM medium containing 10% FBS, 1% PS, 50 ng/mL ascorbic acid, 10 mmol/mL β-glycerophosphate sodium and 4 ng/mL dexamethasone) was added. And the solution was changed every 3 d, ALP staining was performed until the 4th day of culture, and alizarin red staining was conducted on the 14th day to observe the osteogenic differentiation.
Adipogenic differentiation induction was performed on hPDLSCs. After the cells were completely grown, adipogenic induction solution A (α-MEM medium containing 10% FBS, 1% PS, 10 μg/mL insulin, 1 μmol/L indomethacin, and 0.5 mmol/L IBMX) was added. After 3 d, B solution (α-MEM medium containing 10% FBS, 1% PS, and 10 μg/mL insulin) took the place of solution A. One day later, solution A substituted for solution B, while 3 d later, solution B again replaced A. Eventually, oil red O staining was utilized 1 d later for adipogenesis detection.

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Publication 2023

1-Methyl-3-isobutylxanthine Adipogenesis Ascorbic Acid beta-glycerol phosphate Cells Dexamethasone Indomethacin Insulin Osteogenesis Physiologic Calcification Sodium

Triamcinolone Impacts BM-MSC Characteristics

Impact of TP at 10^-8 M concentration on the characteristic features of BM-MSCs were determined by investigating cell surface markers and by evaluating BM-MSCs’ differentiation capacities. For immunophenotyping, 2x10⁵ BM-MSCs were seeded into 6-well tissue culture plates as triplicates, incubated in the presence of 10^-8 M TP for 24 hours followed by collecting cells with trypsinization and labeling with antibodies abovementioned. Cells were immediately read with Beckman Coulter DxFLEX flow cytometry system and analysis was performed on CytExpert software. For evaluation of BM-MSCs’ differentiation capacities, 1x10⁴ BM-MSCs were seeded into 96-well cell culture plates and once they reached 80% confluency, differentiation was initiated by commercial chondrogenesis (Thermo Fisher Scientific, #A1007101), osteogenesis (Thermo Fisher Scientific #A1007201), and adipogenesis (Thermo Fisher Scientific, #A1007001) kits, either supplemented with 10^-8 M TP or not. Differentiation media were replaced twice a week. On the 21^st day of differentiation, cells were fixed with 10% neutral-buffered formalin solution. Chondrogenesis, osteogenesis, and adipogenesis were evaluated by Alcian Blue, Alizarin Red, and Oil Red-O staining, respectively.

Aru B., Akdeniz T., Dağdeviren H., Gürel G, & Yanıkkaya Demirel G. (2023). Testosterone Propionate Promotes Proliferation and Viability of Bone Marrow Mesenchymal Stem Cells while Preserving Their Characteristics and Inducing Their Anti-Cancer Efficacy. Balkan Medical Journal, 40(2), 117-123.

Publication 2023

Adipogenesis Alcian Blue Antibodies Cell Culture Techniques Cells Chondrogenesis Flow Cytometry Formalin Osteogenesis Tissues

Top products related to «Adipogenesis»

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Dexamethasone is a synthetic glucocorticoid medication used in a variety of medical applications. It is primarily used as an anti-inflammatory and immunosuppressant agent.

Oil red o by Merck Group

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Oil Red O is a fat-soluble dye used in histology and cell biology for the staining of neutral lipids, such as triglycerides and cholesterol esters. It is a useful tool for the identification and visualization of lipid-rich structures in cells and tissues.

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Insulin is a lab equipment product designed to measure and analyze insulin levels. It provides accurate and reliable results for research and diagnostic purposes.

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

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Isobutylmethylxanthine is a laboratory reagent used primarily as a phosphodiesterase inhibitor in cell culture and biochemical research applications. It is a synthetic compound that can modulate the activity of cyclic nucleotide signaling pathways. The core function of Isobutylmethylxanthine is to inhibit the breakdown of cyclic AMP and cyclic GMP, which can lead to increased levels of these important signaling molecules in cells.

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Alizarin Red S is a chemical compound used as a dye and a stain in laboratory procedures. It is a red-orange powder that is soluble in water and alcohol. Alizarin Red S is commonly used to stain calcium deposits in histological samples, such as bone and cartilage.

Alizarin red by Merck Group

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Alizarin Red is a laboratory reagent used for the detection and quantitative analysis of calcium in various samples. It is a bright red organic compound that forms a complex with calcium ions, resulting in a distinctive red-colored product. Alizarin Red is commonly employed in histochemical and biochemical applications to stain calcium-containing structures.

What are the key steps involved in the process of adipogenesis?

Adipogenesis is a complex multistep process that involves the differentiation of preadipocytes into mature adipocytes. The main steps include: 1) Growth arrest and clonal expansion of preadipocytes, 2) Activation of key transcription factors like PPAR-gamma and C/EBP-alpha, 3) Upregulation of genes involved in lipid metabolism and adipocyte-specific gene expression, 4) Accumulation of lipid droplets, and 5) Acquisition of the characteristic morphology and functions of adipocytes.

What are some common in vitro and in vivo models used to study adipogenesis?

Researchers utilize a variety of cell culture and animal models to investigate the mechanisms of adipogenesis. Common in vitro models include 3T3-L1 and 3T3-F442A mouse preadipocyte cell lines, as well as primary preadipocytes isolated from rodent or human adipose tissue. In vivo, murine models like diet-induced obesity and genetic knockout/knockin mice are frequently used to study adipogenesis and its regulation in a physiologically relevant context.

How can PubCompare.ai help optimize adipogenesis research?

PubCompare.ai's AI-driven platform can greatly enhance adipogenesis research by helping researchers more efficiently screen and compare protocls from the literature. The tool's advanced AI analysis can pinpooint critical insights, enabling you to identify the most effective protocols for your specific research goals related to adipogenesis. This can improve reproducibility and accuracy, ultimately leading to better research outcomes.

What are some common challenges in studying adipogenesis and how can PubCompare.ai address them?

One key challenge in adipogenesis research is the variability in protocols and methods used across different studies. This can make it difficult to reproduce findings and compare results. PubCompare.ai's AI-driven platform can address this by quickly scanning the literature and highlighting the key differences between protocols, allowing researchers to easily identify the most robust and effective approaches. This can save time and improve the quality of adipogensis research.

Are there different types or variations of adipogenesis that researchers should be aware of?

Yes, there are several variations and subtypes of adipogenesis that have been identified. For example, beige or brite adipogenesis refers to the process of converting white adipocytes into energy-burning beige/brite adipcoytes, which is of great interest for its potential therapeutic applications. Additionally, some researchers have described depot-specific differences in adipogenesis between visceral and subcutaneous fat tissue. Understanding these nuances is important for designing adipogenesis studies that accurately reflect the biological complexities.

More about "Adipogenesis"

Adipogenesis is the process of cellular differentiation where preadipocytes transform into mature adipocytes, or fat cells.
This critical process involves the accumulation of lipid droplets, changes in gene expression and cellular morphology, and the acquisition of the characteristic features of adipocytes.
Adipogenesis is a key aspect of adipose tissue development and is regulated by a complex network of transcription factors, signaling pathways, and epigenetic modifications.
Understanding the mechanisms of adipogenesis is crucial for studying obesity, diabetes, and other metabolic disorders.
Researchers often utilize in vitro and in vivo models to investigate the molecular and cellular events underlying adipocyte differentiation and function.
Common experimental techniques used to study adipogenesis include the use of dexamethasone, Oil Red O staining, indomethacin, insulin, fetal bovine serum (FBS), β-glycerophosphate, 3-isobutyl-1-methylxanthine (IBMX), and Alizarin Red S staining.
Dexamethasone is a synthetic glucocorticoid that plays a key role in adipocyte differentiation, while Oil Red O is a stain used to visualize lipid droplets in adipocytes.
Indomethacin is a non-steroidal anti-inflammatory drug that can also promote adipogenesis.
Insulin is a critical hormone that drives the differentiation of preadipocytes into mature adipocytes, and FBS is a common supplement used in cell culture media to support adipocyte development. β-glycerophosphate is often used as a source of phosphate for the mineralization of adipocytes, and IBMX is a phosphodiesterase inhibitor that enhances adipocyte differentiation.
Alizarin Red S is a stain used to detect the presence of calcium deposits, which can be associated with adipocyte maturation.
By understanding the complexities of adipogenesis and utilizing these experimental tools, researchers can gain valuable insights into the underlying mechanisms of adipose tissue development and the pathogenesis of metabolic disorders.
This knowledge can inform the development of new therapies and interventions to address these pressing health challenges.