3T3-L1 Cells-based In Vitro Screening Service
Your Trusted 3T3-L1 Cells-based In Vitro Screening Partner
CD BioGlyco has a professional Glycobiology Disease Model Development team to provide efficient In Vitro Glycobiology Disease Model Screening services. To effectively study the signaling and mechanisms of glycobiology disease, it is crucial to elucidate its effects at the cellular and molecular levels. The 3T3-L1 cell line is a mature white adipocyte cell model in vitro. Our lab provides high-quality 3T3-L1 cells-based in vitro screening services including stable cell line generation, gene expression analysis, cell signaling analysis, ligand binding test, glyco-biomarkers, and glyco-genes screening.
- Cell culture and optimizing for differentiation
- Our researchers have well-established operational experience in culturing and inducing differentiation of 3T3-L1 cells. To optimize the concentration of differentiation components for 3T3-L1 differentiation, our staff controls the concentration of the culture medium by e.g., increasing the compound content (such as Methyl Isobutyl xanthine, Rosiglitazone), increasing the concentration of Insulin/Dexamethasone, or increasing the concentrations of both Insulin and Dexamethasone.
- Cell labeling and imaging: Our lab provides reliable cell labeling service in labeling buffer. We provide advanced imaging services by spinning disk confocal microscopes.
- Glyco-gene screening and glycosyltransferase assessment
- Gene expression: Our lab uses a real-time polymerase chain reaction (RT-PCR) system to detect and analyze gene expression. Notably, we analyze gene expression using the ∆∆Ct method with multiple housekeeping genes (PPIA, B2M, HPRT).
- Glycosyltransferase analysis: Our researchers determine the strength of different glycosyltransferases by obtaining an inverted mask that segmented the cytoplasmic regions of fat cells.
- Protein extraction and quantification
- CD BioGlyco provides extraction, purification, characterization, and quantification of secreted proteins. We use bicinchoninic acid (BCA) protein assay to determine the protein concentration of whole cell lysates from different samples of 3T3-L1 with different days of differentiation, and then quantify the protein concentration of the standards and samples using a 562 nm laser on an enzyme marker.
- Special analysis
- Importantly, we offer the following high-quality analysis services including MTT assay, SDS-polyacrylamide gel electrophoresis, and immunoblotting.
- We offer a wide range of biomarker analyses based on our clients' needs including but not limited to adiponectin levels, phosphorylated protein kinase B (AKT) and insulin receptor substrate-1 (IRS-1), total insulin receptor, glucose uptake, insulin-like growth factor binding-proteisn-3 (IGFBP-3), IGFBP-6, macrophage colony-stimulating factor (M-CSF), resistin, lipocalin-2, and vascular endothelial growth factor (VEGF).
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Publication
Technology: Cell labelling, Spinning disk confocal microscope, RT-PCR
Journal: Adipocyte
Published: 2020
IF: 3.3
Results: In this article, researchers used 3T3-L1 and human Simpson-Golabi-Behmel syndrome (SGBS) cells to assess the in vitro effects of high doses of insulin. Insulin-treated 3T3-L1 had increased basal glucose internalization and lipid accumulation, which correlated with increased expression of glucose transporter 1 (GLUT1) and fatty acid synthase (FASN). Moreover, the researchers analyzed the expression of selected genes involved in adipocyte differentiation and function. Cebpa and Lep genes had large changes in 3T3-L1 and IL1B in SGBS. Notably, adiponectin were unchanged in 3T3-L1 and SGBS supernatants.
Fig.1 Gene expression in 3T3-L1 and SGBS. (Rossi, et al., 2020)
Applications
- 3T3-L1 can be used for the development and analysis of in vitro insulin-induced insulin resistance models.
- 3T3-L1 cells-based in vitro screening plays a key role in the study of pathogenesis and pathways in diseases such as diabetes, obesity diseases, and insulin resistance.
- 3T3-L1 cells-based in vitro screening can be used to explore the signals generated by adipocytes interacting with target organs.
- 3T3-L1 cells-based in vitro screening can be used to explore the expression of regulatory proteins associated with adipocyte development and maintenance in preadipocyte lineages.
- 3T3-L1 cells-based in vitro screening can be used to develop specific target drugs involved in the pathogenesis of insulin resistance in adipocytes.
Advantages of Us
- Our highly trained glycobiology team provides a one-stop service for the design, development, screening, and data analysis of glycobiological disease models according to our clients' needs.
- Due to the low inter-protein variation, the BCA protein quantification we offer allows for a more accurate determination of the protein concentration of unknown samples.
- Our talented team has a strong background in glycobiology and skillful in vitro screening experience to ensure high-quality deliveries.
With high-quality in vitro cellular models, CD BioGlyco provides our clients with high-throughput target screening services such as glycoprotein, glycogene, and glycobiomarker screening. Our staff is always enthusiastic and professional to serve every client. Please feel free to contact us.
References
- Yudhani, R.D.; et al. In vitro insulin resistance model: a recent update. Journal of obesity. 2023.
- Rossi, A.; et al. In vitro characterization of the effects of chronic insulin stimulation in mouse 3T3-L1 and human SGBS adipocytes. Adipocyte. 2020, 9(1): 415-426.
This service is for Research Use Only, not intended for any clinical use.