3t3 cells adipocytes. The characteristic of this cell line is its ability to differentiate into mature adipocytes upon stimulation with insulin, dexamethasone and methylisobutylxantine when maintained in fetal bovine serum. The cells presently available for research purposes all have unique advantages and disadvantages that one should be aware of when selecting cells. 3T3-L1 adipocytes behave like primary adipocytes in many aspects and provide an excellent model system to study insulin action and signaling (7, 8). Oct 20, 2005 · Much knowledge of adipocyte biology has been learned from cell culture models, most notably 3T3-L1 cells. Jan 2, 2024 · Live cell FRET imaging for LC-acyl-CoAs in 3T3-L1 adipocytes was performed at 37 °C on a laser scanning microscope (Leica SP8) equipped with dual detectors and with continuous Adaptive Focus May 22, 2012 · In the 3T3-L1 cell culture model, temporary exposure of the cells to a combination of insulin, glucocorticoid and an inducer of cAMP signaling triggers adipogenesis, changing the expression of hundreds of structural genes and a variety of transcription factors [3]. The schematic of the differentiation of 3T3-L1 adipocytes. 3T3-L1 adipocytes are the most preferred cell line to study insulin-stimulated glucose uptake, as most of the available muscle cell lines are not insulin sensitive in terms of glucose transport. However, despite its popularity, differentiation of this cell line has been reported to be inconsistent with low efficiency. We would like to manage customer expectations with regard to the potential of the current 3T3 cell line stocks to Feb 8, 2017 · ATCC 3T3-L1, Zenbio 3T3-L1, and OP9 cells were differentiated as described in Methods and assessed for adipocyte differentiation (Nile Red staining of lipid accumulation) and cell proliferation Jan 25, 2021 · We conclude that ColIII is required for 3T3-L1 preadipocyte adipogenesis as well as the formation of actin stress fibres. Apr 14, 2025 · Methods: Immortalized murine embryonic fibroblasts (3T3-L1) cells were differentiated for up to 17 days using a differentiation medium (DM) and a maintenance medium (MM) with or without RGZ or ISO to obtain both the mature white and the beige adipocyte phenotype. The 3T3-L1 cell line is used as an adipocyte differentiation model for the analysis of genes specifically expressed during the differentiation course. Apr 4, 2023 · Aggregating in vitro-grown adipocytes to produce macroscale cell-cultured fat tissue with tunable lipid compositions for food applications As candidates with strong correlation to the adipogenesis timeline we identified CD44 and the acetylation sites FASN_K673 and IDH_K272. We demonstrate that quiescent, fully differentiated 3T3-L1 adipocytes as well as 3T3-L1 preadipocytes can be efficiently transduced with HIV-1-derived lentiviral vectors. Understanding Adipocyte Differentiation. Howard Green of Harvard Medical School, the 3T3-L1 system has been pivotal in advancing the understanding of basic cellular mechanisms associated 3T3 L1 and 3T3 F442A are clonal sublines isolated from 3T3 mouse embryonic fibroblasts, and can be differentiated to adipocytes. Sep 27, 2022 · The results show that only adipocyte-committed 3T3-L1 cells are permissive for the expression of early and late viral mRNAs, as well as viral DNA replication and progeny production, supporting productive HAdV-D36 viral replication, indicating that a greater effect on adipogenesis occurs in adipocytes that support productive viral replication. . 2 T3 is frequently used during brown/beige adipocyte differentiation. Adipose tissue growth involves an increase in adipocyte size and the formation of new adipocytes from precursor cells. Due to their potential to differentiate from fibroblasts to adipocytes, 3T3-L1 cells are widely used for studying adipogenesis and the biochemistry of adipocytes. Hemin enhances the adipose differentiation of mouse 3T3-F442A cells in a medium containing high levels of adipogenic factor that is present in fetal calf serum or calf serum. When injected into mice, 3T3-L1 preadipocytes differentiate and form fat pads that are indistinguishable from normal adipose tissue (Green and Kehinde 1979). Howard Green of Harvard Medical School, the 3T3-L1 system has been pivotal in advancing the understanding of basic cellular mechanisms associated with diabetes, obesity and related disorders. White, beige, and brown adipocytes differ in their handling of lipids and thermogenic capacity, promoting differences in size and morphology. 3T3 L1 is a continuous strain of 3T3 developed through clonal isolation. In summary, the described cell model depicts pathophysiologically hypertrophic 3T3-L1 adipocytes. We have investigated the capacity of OP9 cells, a line of bone marrow-derived mouse stromal cells, to recapitulate adipogenesis. This protocol outlines how to chemically induce the differentiation of 3T3-L1 cells into adipocyte-like cells, and is adapted from the original protocol by Reed & Lane (1980). Moreover, adipocytes release lipids and Of the many in vitro models of adipose tissue [5], the 3T3-L1 fibroblast cell line is one of the most commonly used in vitro models to study adipocyte differentiation and biology because there are well-established protocols for inducing the fibroblasts to become adipocytes which synthesize and accumulate triglyceride [6]. Jul 20, 2023 · ABSTRACT Normalization is a crucial step in gene expression analysis to avoid misinterpretation. To investigate the effect of media height during adipocyte Due to their potential to differentiate from fibroblasts to adipocytes, 3T3-L1 cells are widely used for studying adipogenesis and the biochemistry of adipocytes. Jun 6, 2024 · Highlights Differentiation of 3T3-L1 preadipocytes to mature adipocytes Optimization and validation of transfection methods and reagents for 3T3-L1 siRNA-mediated knockdown strategies in 3T3-L1 preadipocytes and mature adipocytes Summary 3T3-L1 is a model cell line which can be differentiated from preadipocytes into mature adipocytes. 3T3 are murine cells of an established heteroploid cellular line. Established cell lines, such as 3T3-L1 cells, are easier and less Jun 3, 2025 · In summary, the described cell model depicts pathophysiologically hypertrophic 3T3-L1 adipocytes. Because of its potential to differentiate from fibroblasts to adipocytes, the cell line has widely been used in more than 5000 published articles on adipogenesis and the biochemistry of adipocytes [2]. Feed cells every 2-3 days using 3T3-L1 Adipocyte Maintenance Medium until ready for assay. siRNA and cell transfection The 3T3-L1 adipocytes were transfected with siRNA using a lipid-based transfection method (17). ) and were maintained in an incubator set up to 37°C and 5% of CO 2. While Oct 17, 2024 · In 72 h treated adipocytes, no significant differences were found in the viability, glucose uptake or insulin-sensitive and inflammatory gene expression. We first Jul 12, 2025 · Because of its capability to differentiate into adipocytes in vitro, the murine preadipocyte 3T3-L1 cell line is most commonly used to investigate the development of adipocytes. We used geNorm, NormFinder, BestKeeper, RefFinder, and the ∆Ct method to evaluate Download scientific diagram | 3T3-L1 adipocyte differentiation and gene expression validation. Cells can be induced to become adipose-like using the method described in the subculture routine information. The cells were cultured in a serum -containing medium and demonstrated a fibroblast -like morphology. This cell line can be used to study the basic cellular mechanisms associated with diabetes, obesity, and related disorders. Previously, the classical differentiation protocol was optimized by adding the PPARG agonist rosiglitazone. Oct 1, 2005 · As part of our contribution to the Nuclear Receptor Signaling Atlas (NURSA) , we used quantitative real-time PCR to profile the temporal expression of all 49 members of the mouse nuclear receptor superfamily at selected time points during differentiation of 3T3-L1 cells into mature, lipid-bearing adipocytes using two differentiation inducers Abstract In vitro models have been invaluable in determining the mechanisms involved in adipocyte proliferation, differentiation, adipokine secretion and gene/protein expression. Plate the cells in proliferation media (8% CS/DMEM) on treated polysterene culture dishes from Corning and incubate them at 37 ˚C in 10% CO2. 3T3-L1 adipocytes are not a good model of adipocytes for canonical catecholamine plasma membrane transporters, and caution should be used when applying the The 3T3-L1 cell line is one of the most well-characterized and reliable models for studying the conversion of preadipocytes into adipocytes. The phenotype of Col 3a1 −/− 3T3-L1 cells is very similar to that of Gpr 56 −/− 3T3-L1 cells, suggesting a functional relationship between ColIII and Gpr56 in preadipocytes. Originally derived from Swiss mouse embryo tissue by Dr. The 3T3-L1 cell line is a well-characterized and reliable model for studying preadipocyte conversion into adipocytes. The 3T3-L1 cell line, derived from 3T3 cells, is widely used in biological research on adipose tissue. These cell lines are commonly used a model to study fat metabolism. In Glass microfluidic devices have been fabricated to monitor the secretion of glycerol or fatty acids from cultured murine 3T3-L1 adipocytes. In addition to classical white and brown adipocytes that are derived from different cell lineages, beige adipocytes have also been identified, which have characteristics of both white and brown adipocytes. Visualizing other cellular compartments contributing to the cellular machinery requires additional cell culturing for Jun 19, 2015 · Abstract Differentiated 3T3-L1 adipocytes are a widely used in vitro model of white adipocytes. 3T3-L1 3T3-L1 cells have been widely used as a model for adipogenesis. 2 Capsaicin . Highlights: After differentiation, these cells accumulate large amounts of triglyceride fat Triglyceride fat accumulation can be reduced by lipolytic agents Express insulin receptor 3T3 L1 The 3T3-L1 cell line is commonly used to study adipocyte biology, adipocyte cell differentiation, and related cellular and molecular mechanisms. The 3T3-L1 cell is a model cell line, which could be inducted from preadipocytes (fibroblast-like cells) to the differentiated adipocytes (round cells with lipid droplets) (Figure 1). INTRODUCTION 3T3-L1 adipocytes have been fundamental in metabolic disease research for 30 years. Most studies investigate such mechanisms using mature adipocytes that have been chemically induced to differentiate for 7 days in media containing 25 mM glucose. Adipocytes are specialized cells with pleiotropic roles in physiology and pathology. One of the main cellular systems used to study the underlying physiological and biological processes is the 3T3-L1 preadipocyte differentiation model. When 3T3-L1 cells in fetal bovine serum are treated with dexamethasone alone, a supply of an Jun 1, 2012 · Abstract In this note, we present a detailed procedure for highly effective and reproducible 3T3-L1 cell differentiation. In the current studies, adipocytes are perfused in a reversibly-sealed cell chamber and secreted products are Mar 1, 2019 · The differentiation efficiency of 3T3-L1 preadipocytes is an essential factor affecting studies on cellular mechanisms associated with obesity, diabetes, and related disorders. In the present study, 3T3-L1 cells from the DI group exhibited morphological changes to adipocyte-like structures, with lipid droplet accumulation observed on days 5 and 10 after adipogenic induction. Adipocytes play a vital role in glucose metabolism. The pre-adipocyte 3T3-L1 cell culture line, derived from mouse embryos, is a widely used model for the study of adipocyte differentiation. Methodology/Principal Findings 2. Oct 8, 2024 · Firstly, only one cell line, preadipocyte cell line, 3T3-L1, was used in this work, although this is the most extensively studied cell line in this adipocyte-related research field. While 3T3-L1 preadipose cells derived from mouse embryo are the most commonly used cell line for adipose research [7], the human SGBS pre-adipocytes, originally derived from adipose tissue obtained from a patient affected by Simpson-Golabi-Behmel syndrome (SGBS) [8], have been used only in recent years. 3T3 L1 pre adipocytes after differentiation to adipocytes serve as excellent in vitro models and are useful tools in understanding the glucose metabolism. While results generally aligned between SGBS and 3T3-L1 cells, details appeared cell line specific. Researchers also use this cell model to study obesity and insulin resistance. 3T3-L1 is one of the most commonly used adipocyte cell lines. Lipid accumulation and adipokine secretion profiles were analyzed. The 3T3-L1 cell line is commonly used to study adipocyte biology, adipocyte cell differentiation, and related cellular and molecular mechanisms. Some clones of this cellular line, when cultured under adequate conditions differentiate into adipocytes. Incubate for 2-3 days. Therefore, the molecular mechanism of adipogenesis is intensively studied in vitro using different preadipocyte clonal cell lines from mice or rats. This model can be used for knockdown, overexpression, and stimulation studies, thereby serving as an alternative to primary cells isolated from DIO mice. 26 Prolonged Rosiglitazone and T3 in addition to IBMX also increased beige‐like gene profiles in 3T3‐L1 adipocytes. Sep 16, 2020 · The 3T3-L1 cell is a model cell line, which could be inducted from preadipocytes (fibroblast-like cells) to the differentiated adipocytes (round cells with lipid droplets) (Figure 1). Keywords: 3T3-L1 cells; Adipogenesis; Cell culture; Intracellular lipid accumulation; Lipid staining. Furthermore, these adipocytes showed significantly reduced insulin sensitivity, adiponectin secretion, and lipolytic enzyme expression, accompanied by increased IL6 and leptin secretion. Aug 2, 1991 · Mammalian 3T3-L1 cells differentiate into adipocytes after continuous exposure to pharmacological doses of insulin or physiological doses of insulin-like growth factor I (IGF-1). Hemin also promotes adipose differentiation in a medium containing 10% cat serum, which has very little adipogenic factor; this effect is in contrast to that The 3T3-L1 cell-line is a widely used model system for the analysis of adipocyte differentiation. When OP9 Differentiation of 3T3-L1 cells into adipocyte-like cells This protocol outlines how to chemically induce the differentiation of 3T3-L1 cells into adipocyte-like cells. Adipogenesis is a complex process influenced by various cellular interactions within adipose tissue, which plays a critical role in metabolic homeostasis. Differentiation of multi-potential 3T3-L1 fibroblasts into lipid laden adipocyte-like cells is one of the most common in vitro models used in the study of adipocyte biology. A widely used histological and immunohistochemical staining and mass spectrometry lipidomics are mainly aimed for examining lipid droplets (LDs). Gregoire, Francine M. Culturing cells on bio-gels are believed to provide a more in vivo-like extracellular matrix. 78: 783–809, 1998. 3T3-L1 differentiation is an economical and convenient way to generate adipocyte-like cells for experiments. Here, we present a protocol for changing gene expression in 3T3-L1 materials from at least two different distribution lots were tested for their ability to differentiate in response to a mixture of insulin, dexamethasone, and IBMX (methylisobutylxanthine). The cells are not contact inhibited. When 3T3-L1 cells in fetal bovine serum are treated with dexamethasone alone, a supply of an Incubate for 3 days. Apr 1, 2025 · In this regard, we investigated the role of IMPDH, a vital enzyme in cell viability, and its interaction with ARL13B, a major cilium protein, on the cilia development among 3T3-L1 cells while undergoing differentiation to lipid-accumulating adipocytes. Here, we present a protocol for combining qualitative and quantitative analysis of lipid droplets in mature 3T3-L1 adipocytes using oil red O. While The 3T3-L1 murine cell line is a robust and widely used model for the study of adipogenesis and processes occurring in mature adipocytes. We surveyed the literature, the gene expression omnibus and the sequence Abstract Background 3T3-L1 cells are widely used to study adipogenesis and insulin response. Rev. We then detail The 3T3-L1 cell-line is a widely used model system for the analysis of adipocyte differentiation. Scale bar: 100 μm. To this end, therefore, this experimental work aims to define the aspects listed above in order to establish an adipocyte differentiation protocol using 3T3-L1 cells for the investigation of the modulation of the de novo browning process and the molecular targets involved. 3T3-L1 cells cultured on Matrigel® significantly alteregd their proliferation and differentiation as compared to growth on tissue culture-coated Apr 18, 2015 · Abstract Differentiated 3T3-L1 adipocytes are a widely used in vitro model of white adipocytes. The differentiation of 3T3-F442A cells is accompa- nied by changes in cell morphology, 1982). Expressing exogenous genes in 3T3-L1 cells can be challenging. Smas, and Hei Sook Sul. However, the dysfunctional characteristics commonly observed in obesity including adipocyte Dec 28, 2019 · Background Adipogenesis is the process of adipocytes formation from unspecialized progenitor cells called mesenchymal stromal cells. Jun 1, 2012 · The preadipose cell line 3T3-L1 was originally developed by clonal expansion from murine Swiss 3T3 cells [1]. Protein concentration in cell lysates was determined using BCA TM protein assay (Thermo Scientific). Jun 10, 2023 · Real-time metabolic analysis revealed that the metabolic capacity shifted from glycolysis to mitochondrial respiration in differentiated 3T3-L1 cells in the 3D culture condition but not in those in the 2D cultured condition, suggesting that adipogenic differentiation in 3D culture induces a metabolic phenotype of well-differentiated adipocytes. Here, we describe methods used for the culture and transformation of these preadipocytes into mature adipocytes and quantification of intracellular lipid accumulation using the lipid specific dye, Oil Red O. The representative phase contrast (B) and oil red O staining (C) Images of 3T3-L1 cells at the preadipocyte, 4-day-differentiated, and 8-day-differentiated adipocyte stages. Apr 1, 2014 · We used the 3T3-L1 cell differentialtion model to prepare nondifferentiated (preadipocytes) and differentiated cells (adipocytes), and performed customized PCR arrays containing genes that encode mouse chemokines and chemokine receptors. In 1971, Green and Kehinde established a subline of 3T3 cells, called 3T3-L1, which demonstrated a greater propensity for differentiation into adipocytes and a higher affinity for lipid deposition. Moreover, adipocytes release lipids and Oct 17, 2024 · Of the many in vitro models of adipose tissue [5], the 3T3-L1 fibroblast cell line is one of the most commonly used in vitro models to study adipocyte differentiation and biology because there are well-established protocols for inducing the fibroblasts to become adipocytes which synthesize and accumulate triglyceride [6]. This cell model has several applications in obesity and insulin resistance research. Expression of transfected ras oncogenes led to differentiation of Nov 15, 2024 · Adipocytes derived from 3T3-L1 cells are a gold standard for analyses of adipogenesis processes and the metabolism of fat cells. , Cynthia M. Differentiation of 3T3-L1 MBX cells was at first optimized and then maintained in the presence of fatty acids cocktail combination to induce the obese condition. It is important to feed the preadipocytes every couple of days to avoid letting them get too confluent (>70%), if you want to continue to passage them and differentiate them at a later date. This study provides a modified approach to in vitro odeling adipocyte inflammation and insulin resistance. Remove the 3T3-L1 Differentiation Medium and replace with 3T3-L1 Adipocyte Maintenance Medium. (A) Induction of 3T3-L1 adipocyte differentiation by DEX and IBMX initially results in cell growth. Material and Methods The 3T3-L1 cell line after differentiation was used as an adipocyte model. The 3T3-L1 pre-adipocyte cell line is widely used to study the fat cell differentiation in vitro. Here, we present a protocol for changing gene expression in 3T3-L1 (pre)adipocytes using small interfering RNA (siRNA)-mediated knockdown. Here we show that 3T3-L1 adipocytes display features of multiple adipocytes lineages. Keywords: 3T3-L1 cells; Adipocyte; in vitro model; inflammation; insulin resistance. In the present report, we have interfered with changes in actin assembly and cell morphology using the decreased syn- thesis and assembly of cytoskeletal proteins, and extracellular matrix protein fibronectin. This study aimed to develop a novel in vitro three-dimensional (3D) co-culture model using murine 3T3-L1 pre-adipocytes, J774 macrophages, and NIH-3T3 fibroblasts to investigate adipogenic differentiation and inflammatory pathways. (D) The quantitative results of oil red O May 26, 2016 · The differentiation of adipocytes is tightly regulated by a variety of intrinsic molecules and also by extrinsic molecules produced by adjacent cells. When injected into mice, 3T3-L1 preadipocytes differentiate and form fat pads that are indistinguishable from normal adipose tissue (Green and Kehinde 1979 Apr 3, 2025 · The TULP family was first identified in progressively obese mice, and TULP3, as a member of its family, has been much studied in tumor cells, but studies on its role in adipocytes have not yet been reported. Jun 20, 2024 · Summary By differentiating into mature adipocytes, 3T3-L1 cells can be utilized as a model cell line to investigate (pre)adipocyte function in vitro. In this report, we evaluated the use of lentivirus-mediated gene transfer into 3T3-L1 mature adipocytes. Dietary exposures occurring over a specific period of time might cause long-lasting and even permanent changes in gene expression regulated by epigenetic mechanisms 3T3-L1 is a fibroblast that was isolated from the embryo of a mouse. Our datasets on SGBS and 3T3-L1 adipogenesis dynamics are accessible for further mining. The assay was established according to published protocols and is relatively easy to perform. The ability of 3T3‐L1 cells to differentiate to beige adipocytes was previously shown using capsaicin, which induced a ‘brite’ phenotype. Appearance of adipocytes can take weeks to achieve. We built a Sep 3, 2023 · The 3T3-L1 murine adipocyte cell line remains one of the most widely used models to study the mechanisms of obesity and related pathologies. This enhancement is more prominent in the presence of insulin. 3T3-L1 Summary 3T3-L1 is a model cell line which can be differentiated from preadipocytes into mature adipocytes. This work tries to establish and characterize an alternative model of cultured adipocytes that is easier to work with than the 3T3-L1 cells. 3T3-L1 preadipocytes lend themselves very well to the study of adipogenesis. Jun 21, 2024 · Here, we present a protocol for changing gene expression in 3T3-L1 (pre)adipocytes using small interfering RNA (siRNA)-mediated knockdown. The fibroblastic like cells can be induced by hormones to differentiate into mature adipocytes. Their adipogenic potential decreases with time in the culture. The differentiation of 3T3-L1 cells into adipocytes is closely mimics the in vivo differentiation pathway of adipocytes. The 3T3-L1 model has several limitations, including the requirement of 2 weeks to generate adipocytes and the waning of adipogenic potential in culture. Aug 9, 2023 · Here, we describe 3T3-L1 MBX cells as a culture model for studying obese adipocytes and their effects. However, 3T3-L1 cells after freezing and thawing for many To learn more about the expression and regulation of CDO and ADO in adipocytes, we examined the levels of taurine biosynthetic enzymes in rat adipose tissues and in differentiating 3T3-L1 cells and their responses to cysteine concentration. Cells were treated with Isoproterenol (ISO), ANGPTL3-Fld, or a combination of both. Dec 1, 2000 · The 3T3-L1 cell line is one of the most well-characterized and reliable models for studying the conversion of preadipocytes into adipocytes. In this study, Oct 1, 2023 · Real-time metabolic analysis revealed that the metabolic capacity shifted from glycolysis to mitochondrial respiration in differentiated 3T3-L1 cells in the 3D culture condition but not in those in the 2D cultured condition, suggesting that adipogenic differentiation in 3D culture induces a metabolic phenotype of well-differentiated adipocytes. The traditional approaches adopted in pre adipocyte differentiation are lengthy exercises inv … The differences between 3T3-L1 adipocytes and rat mesenteric PVAT/adipocytes suggest that the cells neighboring adipocytes in PVAT may play a large role in the function of these adipocytes. Several types of fat cells with distinct metabolic properties coexist in various anatomically defined fat depots in mammals. This study found that the expression of TULP3 showed an increasing trend in the differentiation of 3T3-L1 cells, and overexpression of TULP3 enhanced the proliferation and differentiation We would like to show you a description here but the site won’t allow us. However, studies on 3T3-L1 adipocytes are hampered by the fact that genetic modification of mature adipocytes is notoriously difficult. Differentiation of 3T3-L1 cells is commonly induced by an adipogenic cocktail containing insulin, dexamethasone (DEX), and 3-isobutyl-1-methylxanthine (IBMX). During the Another important cell line derived from murine Swiss 3T3 cells is the 3T3-F442A cell line, which displays a more advanced commitment in the adipocyte lineage than 3T3-L1 due to its isolation from a third selection of clones that converts into fat cells clusters of increased size and at a higher frequency. The 3T3-L1 cell line, which is used extensively as an in vitro model to generate adipocyte-like cells, was used as the adipocyte differentiation model. 3T3-L1 adipocyte differentiation was conducted in a Petri dish in vitro. During the process of differentiation, the cells undergo a change from the elongated fibroblastic shape to a round or oval form … Jun 16, 2023 · To minimize the effects of experimental variation, we mixed 3T3-L1 and SGBS cells and used computational analysis to demultiplex transcriptomes of mouse and human cells. For the last 20 years, the cellular and molecular mechanisms of adipocyte Adipocyte cell culture has proven indispensable for the study of the cellular pathophysiology of energy metabolism. The present study indicates that 3T3-L1 cells can differentiate to beige-like adipocytes by prolonged treatment with the mixture of T3, IBMX and Rosi and that the gene expression profile of the adipocytes is distinct from those previously induced from white fat depots. Jun 1, 2012 · In this note, we present a detailed procedure for highly effective and reproducible 3T3-L1 cell differentiation. Mouse 3T3-L1 Cell Line Systems ZenBio now offers Cryopreserved and Cultured 3T3-L1 Preadipocytes! 3T3-L1 murine adipocytes have been fundamental in metabolic disease research for over 30 years. We describe steps to perform the knockdown of a certain gene prior to differentiation (day 4) to analyze the impact on adipogenesis. — The adipocyte plays a critical role in energy balance. Contact inhibition arrests preadipocyte proliferation at confluence. Numerous mechanisms including epigenetic regulation modulate the correct progress of this process. Physiol. Purchase high quality cryopreserved mouse embryonic fibroblast NIH 3T3-L1 cells from ECACC for adipocyte differentiation using insulin, dexamethasone, and IBMX. Oct 28, 2003 · Obesity is associated with many serious afflictions such as cardiovascular disease, cancer, and diabetes. 3T3-L1 pre-adipocytes were cultured in initiated from publication: Systematic characterization of Cell culture and differentiation of 3T3-L1 cells Murine 3T3-L1 cells (American Type Culture Collection, Rockville, MD) were cultured in DMEM containing 25 mM glucose, 10% calf bovine serum (Invitrogen, Carlsbad, CA), and 1% penicillin and streptomycin (Gibco, Invitrogen Corp. Our study revealed that autophagy is specifically required for the initial stage of adipogenic differentiation. This application note determines if 3T3-L1 adipocyte differentiation is dependent on contact inhibition and tests the OP9 cell line as a possible substitute for 3T3-L1 cells in adipocyte assays. ISO was used as the standard β-adrenergic lipolysis mediator. Robust and well-characterized, 3T3-L1 pre-adipocytes differentiate and form fat pads indistinguishable from normal adipose tissue when injected into mice. 3T3-L1 cells have a fibroblast-like morphology, but, under appropriate conditions, they differentiate into an adipocyte-like phenotype. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of 10 candidate housekeeping genes in non-differentiated (ND) and differentiated (DI) 3T3-L1 cells on days 5 and 10. ylhwe djm5 hqlehh ercy eka dndw gda 06n 93cznz gho