Platelet derived growth factor pdgf

Manufactured by R&D Systems

Sourced in United States

Platelet Derived Growth Factor (PDGF) is a laboratory product that functions as a growth factor. It is involved in the regulation of cellular processes such as cell growth, cell differentiation, and cell migration.

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Lab products found in correlation

Ez link sulfo nhs biotin, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Tnf α, by R&D Systems (1 mentions) Interleukin 6 il 6, by R&D Systems (1 mentions) Recombinant transforming growth factor β, by R&D Systems (1 mentions) Anti ki67 antibody, by Abcam (1 mentions) Zymosan a zya, by Merck Group (1 mentions) Imagexpress micro confocal, by Molecular Devices (1 mentions) Recombinant human tgf β1, by R&D Systems (1 mentions) Cyquant nf cell proliferation assay, by Thermo Fisher Scientific (1 mentions)

5 protocols using platelet derived growth factor pdgf

Stimulating NIH3T3 Cells with Growth Factors

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NIH3T3 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, from Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Scientific, Waltham, MA, USA) and maintained in humidified atmosphere of 5% CO₂ at 37^°C. For cell treatments, recombinant transforming growth factor-β (TGF-β) (5 ng/mL), tumor necrosis factor-α (TNF-α) (5 ng/mL), vascular endothelial growth factor (VEGF)-165 (20 ng/mL), platelet-derived growth factor (PDGF) (10 ng/mL) or interleukin-6 (IL-6) (10 ng/mL) (all purchased from R&D Systems, Minneapolis, MN, USA) were added to the medium for 24 hrs. The macrophage cell line RAW264.7 was cultured in DMEM containing 10% FBS as described [48 (link)].

Guo X., Sun M., Dai C., Zhang X., Yin Q., Ling J., Li X., Wu X., Jiang F, & Wang J. (2017). Macrophage-stimulated microRNA expression in mural cells promotes transplantation-induced neointima formation. Oncotarget, 8(18), 30100-30111.

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Zymosan A Activation Assay

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Zymosan A (ZyA) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Cytokines and platelet-derived growth factor (PDGF) were from R&D Systems (Minneapolis, MN, USA). Compound 968, 5-(3-bromo-4-(dimethylamino)phenyl)-2,2-dimethyl-2,3,5,6-tetrahydrobenzo[a]phenanthridin-4(1H)-one, was obtained from Calbiochem (La Jolla, CA, USA). Anti-glutaminase and anti-Ki-67 antibodies were obtained from Abcam (Cambridge, UK).

Takahashi S., Saegusa J., Sendo S., Okano T., Akashi K., Irino Y, & Morinobu A. (2017). Glutaminase 1 plays a key role in the cell growth of fibroblast-like synoviocytes in rheumatoid arthritis. Arthritis Research & Therapy, 19, 76.

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Spatial Mapping of Growth Factor Gradients

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Platelet Derived Growth Factor (PDGF, R&D Biosystems) was biotinylated (NHS Sulfo EZ Link Biotin) according to manufacturer’s protocol (Fisher) to allow for post-fixation identification. MAP particles (no hep, Hep_Low, and Hep_High) were embedded (1:100) in 100μL collagen-agarose gel (1 mg/mL collagen, 1% agarose) for structural support. Particles were labeled with different Alexa Flour maleimide tags to identify the different types. After gelation at 37°C, the gels were soaked in 10% FBS for 8 hours to mimic in situ conditions. Finally, the solutions were removed and then incubated with the growth factor of interest at 1 μg/mL in 1% BSA for 48 hours on a shaker plate at 37°C. Following incubation, the gels were frozen in OCT and then sectioned into 10μm sections using a cryostat. To visualize the gradients, the slides were rehydrated with PBS and then stained with streptavidin labeled with Cy5 (1:200 dilution) for 1 hour at room temperature. Slides were washed three times with PBS and then coverslipped and imaged at 20x using an ImageXpress Micro-Confocal (Molecular Devices).

Pruett L., Jenkins C., Singh N., Catallo K, & Griffin D. (2021). Heparin Microislands in Microporous Annealed Particle Scaffolds for Accelerated Diabetic Wound Healing. Advanced functional materials, 31(35), 2104337.

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Bioactive PDGF Delivery from Heparin Scaffolds

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Platelet Derived Growth Factor (PDGF, R&D Biosystems) was biotinylated (NHS Sulfo EZ Link Biotin) according to manufacturer’s protocol (Fisher) to allow for post-fixation identification. 15μL MAP scaffolds (10% Hep_High) were embedded in 100μL collagen-agarose gel (1 mg/mL collagen, 1% agarose) for structural support. Heparin microgels were labeled with AF555 for post-fixation identification. After gelation at 37°C, the gels were soaked in 10% FBS for 4 hours to mimic in situ conditions. Next, the solutions were removed and gels were incubated with biotinylated PDGF at 1 μg/mL in 1% BSA for either 5 min, 1 hour, 8 hour, or 48 hour (N=3 for each time point) on a shaker plate at 37°C. Following incubation, the gels were fixed with paraformaldehyde for twenty minutes followed by three five-minute PBS washes. Finally, gels were stained with AF647 Streptavidin to visualize growth factor localization for 48 hours, followed by 48 hours of washing in a PBS solution. After PBS washes, the gels were imaged at 20x using a Zeiss 780 Confocal Microscope in the Keck Center for Biologic Imaging.

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Cell Proliferation Assay with PDGF and TGF-β

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Dishes were coated with cell-derived fibronectin (purified from conditioned media of human lung fibroblasts; 20 μg/ml) then rinsed and blocked as described above. Cells were seeded at 5000 cells/well in triplicate wells per condition and incubated with 1 % serum + DMEM in addition to 20 ng/mL of Platelet Derived Growth Factor (PDGF, R&D systems, Minneapolis, MN) or 2 ng/mL of human recombinant TGF-β1 (R&D systems, Minneapolis, MN). The cells were then incubated at 37 °C and 5 % CO₂ for 24 hours. Proliferation was assessed using the CyQuant® NF Cell Proliferation assay (Life Technologies) following the manufacturer’s instructions. Fluorescence intensity was measured with a microplate reader. Blank wells containing dye only and background (cells, no dye) were subtracted from the average emission of three wells for each condition.

Matthes S.A., LaRouere T.J., Horowitz J.C, & White E.S. (2015). Plakoglobin expression in fibroblasts and its role in idiopathic pulmonary fibrosis. BMC Pulmonary Medicine, 15, 140.

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