Fibroblast growth factor (fgf)

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The FGF is a laboratory equipment product from Thermo Fisher Scientific. It is designed for the measurement and analysis of fibroblast growth factors. The core function of the FGF is to provide accurate and reliable data for researchers and scientists working in the field of cellular biology and biochemistry.

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Lab products found in correlation

364 protocols using fibroblast growth factor (fgf)

Directed Differentiation of hiPSCs to Hematopoietic Progenitors

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H1 hESCs were obtained from WiCell Research Institute (Madison, WI). H1 hESC line, the iETS1 H1 hESC line and the tdTomato H1 hESC lines were maintained on matrigel in mTeSR1 medium. Cells were passaged every 3–4 days using 0.5 mM EDTA in PBS. The hESC lines were differentiated on ColIV coating plate (Uenishi et al., 2014 (link)). Singular iETS1 hESCs were plated 10,000 cells/cm² to 6 well plates coated with ColIV (Sigma). After 1 day, media was changed to IF9S media with 50 ng/ml FGF (Peprotech), 50 ng/ml BMP4 (Peprotech), 15 ng/ml Activin A (Peprotech) and 2 mM LiCl (Sigma). On day 2, media was changed to IF9S media with 50 ng/ml FGF and 50 ng/ml VEGF (Peprotech). On day 4, media was changed to IF9S media with 50 ng/ml FGF, 50 ng/ml VEGF, 50 ng/ml TPO (Peprotech), 50 ng/ml SCF (Peprotech), 50 ng/ml IL-6 (Peprotech) and 10 ng/ml IL-3 (Peprotech). On day 6, IF9S media with cytokines the same as day 4 media was added to the culture.

Park M.A., Kumar A., Jung H.S., Uenishi G., Moskvin O.V., Thomson J.A, & Slukvin I.I. (2018). Activation of Arterial Program Drives Development of Definitive Hemogenic Endothelium with Lymphoid Potential. Cell reports, 23(8), 2467-2481.

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Bovine Mammary Epithelial Cell Culture

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PS cells are a bovine mammary epithelial cell line obtained in our laboratory from secretory parenchyma [23 (link)]. Cells were cultured at 37°C in 5% CO₂ in Advanced DMEM/F-12 medium (Gibco) containing 4 ng/mL of hydrocortisone (Gibco), 2mM of glutamine (Gibco), 20 mM of HEPES (Biowhittaker), IGF-I (Insulin-like Growth Factor; 10 ng/mL; Peprotech), FGF (Fibroblast Growth Factor; 5 ng/mL; Peprotech) and EGF (Epidermal Growth Factor; 5 ng/mL; Sigma).

Védrine M., Berthault C., Leroux C., Répérant-Ferter M., Gitton C., Barbey S., Rainard P., Gilbert F.B, & Germon P. (2018). Sensing of Escherichia coli and LPS by mammary epithelial cells is modulated by O-antigen chain and CD14. PLoS ONE, 13(8), e0202664.

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Tumor Sphere Formation Assay

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MIA PaCa-2 and PANC-1 cells were plated in ultra-low attachment 6-well plates (Corning, NY, USA) at a density of 5,000 cells/well. The cells were maintained in serum-free DMEM/F12 supplemented with 2% B27 supplement (Gibco, MA, USA), 1% N2 supplement (Gibco), 20 ng/ml epidermal growth factor, EGF (Gibco), 20 ng/ml fibroblast growth factor, FGF (Gibco), and 1% antibiotic-antimycotic (Invitrogen, CA, USA). The number and size of tumor spheres formed were evaluated by light microscopy after 7-12 days.

Kwon H.M., Kang E.J., Kang K., Kim S.D., Yang K, & Yi J.M. (2017). Combinatorial effects of an epigenetic inhibitor and ionizing radiation contribute to targeted elimination of pancreatic cancer stem cell. Oncotarget, 8(51), 89005-89020.

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Culturing Spry124 Mutant Mouse Embryonic Fibroblasts

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Spry124^fl/fl and Spry124^−/− MEFs were cultured in DMEM containing 10% FBS as previously described (Akbulut et al., 2010 (link)). Unsynchronized MEFs were maintained in DMEM containing 10% FBS for 24 hours. Serum starved MEFs were maintained in DMEM containing 0.1% FBS for 24 hours. FGF-treated MEFs were maintained in DMEM containing 0.1% FBS for 20 hours after which 10 ng/mL FGF (Life Technologies) was directly added to the media for an additional 4 hours, unless otherwise noted. Additional details related to retroviral transduction, inhibitor treatments, siRNA transfection, protein and RNA isolation, and biological assays can be found in the Supplemental Experimental Procedures.

Nabet B., Broin P.Ó., Reyes J.M., Shieh K., Lin C.Y., Will C.M., Popovic R., Ezponda T., Bradner J.E., Golden A.A, & Licht J.D. (2015). Deregulation of the Ras-Erk Signaling Axis Modulates the Enhancer Landscape. Cell reports, 12(8), 1300-1313.

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Glioblastoma Cell Culture Protocols

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Experiments were conducted using the GBM cell lines D54-MG (WHO IV, GBM, gifted by Dr. D.D. Bigner, Duke University, Durham, NC, USA) and U251MG and three patient-derived xenolines labeled GBM12, GBM14, and GBM22. Adherent cell cultures were maintained in DMEM/F12 (ThermoFisher, Waltham, MA, USA, #11320-082) with 7% fetal bovine serum (Aleken Biologicals, Nash, TX, USA). GBM14 and GBM22 were maintained in DMEM (ThermoFisher #21041-025) supplemented with B-27 Supplement without vitamin A, (ThermoFisher #12587010), EGF (ThermoFisher #PHG0311), FGF (ThermoFisher PHG0261), sodium pyruvate (ThermoFisher #11360-070), gentamicin (Fisher BW17 518Z), and amphotericin (Fisher #BP264550). GBM12 cells were maintained in a NeuroBasal (ThermoFisher #12348017) media supplemented with B-27, EGF, FGF, L-glutamine (ThermoFisher #21041-025), amphotericin, and gentamicin. Cultures were incubated at 37 °C and in 10% a CO₂ atmosphere. Brightfield images of cell cultures were captured on a Nikon Eclipse TS100 (Melville, NY, USA) inverted microscope.

Umans R.A., Martin J., Harrigan M.E., Patel D.C., Chaunsali L., Roshandel A., Iyer K., Powell M.D., Oestreich K, & Sontheimer H. (2021). Transcriptional Regulation of Amino Acid Transport in Glioblastoma Multiforme. Cancers, 13(24), 6169.

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Derivation and Differentiation of hiPSC Motor Neurons and Astrocytes

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Control hiPSC motor neuron and cortical neuron progenitors were derived as described [71 (link)] from multiple donors (Table 2). These cells were cultured on Matrigel (Corning) coated plates in base medium, comprised of 50% NeuroBasal (Gibco), 50% advanced DMEM (Gibco), supplemented with B27 and N2 (gibco), 100 μg/ml Pen-Strep (Gibco), and 2 mM L-alanyl-L-glutamine dipeptide (Gibco). For expansion of progenitors, FGF (20 ng/ml) (Gibco) was added to base medium. Differentiation of MN progenitors was achieved using base medium with compound E (Enzo) (0.1 μm) and the growth factors BDNF (10 ng/ml) and GDNF (10 ng/ml) unless stated. Astrocytes were generated from iPSC using a modified protocol described in Hall and colleagues [71 (link)]. Derived astrocyte progenitors were cultured in neuronal base medium with FGF (20 ng/ml) (Gibco) and EGF (20 ng/ml) (Thermo). For differentiation, astrocytes were cultured in base medium without growth factor supplements. All cells were cultured with 5% CO2 in humidified atmosphere at 37°C (Table 3).

Hagemann C., Bailey M.C., Carraro E., Stankevich K.S., Lionello V.M., Khokhar N., Suklai P., Moreno-Gonzalez C., O’Toole K., Konstantinou G., Dix C.L., Joshi S., Giagnorio E., Bergholt M.S., Spicer C.D., Imbert A., Tedesco F.S, & Serio A. (2024). Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs. PLOS Biology, 22(3), e3002503.

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FGF Bioactivity Evaluation via Wound Healing

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To determine FGF biological activity, CytoSelect 24-well Wound Healing Assay (Cell Biolabs, United States) was used. The experiments were performed according to the manufacturer’s protocol. Briefly, inserts of 0.9 mm wound field were placed into previously collagen-coated [50 μg/ml in 0.1 M acetic acid (Millipore, United States)] 24-well plate. 10 × 10⁵ of NIH/3T3 mouse fibroblasts in 500 μl low-glucose DMEM (Sigma, United States) with 10% FBS (Sigma, United States) and 1% of penicillin/streptomycin (Sigma, United States) were incubated at 37°C overnight. The following three groups were used for the experiments: PBS, 50 ng/ml of FGF (Invitrogen, United States), and FGF released from the cryogel (day 3). Images were retrieved at 0 h and then every 2 h using EVOS^TM FL Auto 2 Imaging System (Invitrogen, United States). After 24 h, cells were fixed with 4% formaldehyde and stained with DAPI. Images were obtained and the surface area of the migrated cells was measured using Fiji software.

Jimi S., Jaguparov A., Nurkesh A., Sultankulov B, & Saparov A. (2020). Sequential Delivery of Cryogel Released Growth Factors and Cytokines Accelerates Wound Healing and Improves Tissue Regeneration. Frontiers in Bioengineering and Biotechnology, 8, 345.

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Culturing Murine and Human DIPG Cell Lines

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p53 WT (PDGF-B; H3.3K27M; PTEN deficient ⁵¹) and p53-deficient (PDGF-B; H3.3K27M; p53 loss (Cre), or NS-PKC) ⁵² murine DIPG lines (created in the laboratory of O.J.B.) were cultured in NeuroCult medium with proliferation supplement (STEMCELL Technologies), 100 units/mL penicillin, and 100 μg/mL streptomycin (Invitrogen), 2ug/mL Heparin (Sigma-Aldrich), 20ng/mL FGF (Peprotech) and 10ng/mL EGF (Peprotech). The human p53 deficient DIPG cell line, SF8628 ⁵³ (generated by R. Hashizume, Northwestern University) was cultured in DMEM medium (Gibco-ThermoFisher), supplemented with 10% fetal bovine serum (Seradigm), 100 units/mL penicillin, and 100 μg/mL streptomycin (Invitrogen). The human p53 WT DIPG cell line, SU-DIPG ⁵⁴ (generated by M. Monje, Stanford University) was cultured in Neurobasal media (-A) (Invitrogen), B27 (-A) (Invitrogen), 10ng/mL Heparin (Sigma-Aldrich), 20ng/mL FGF (Peprotech) and 10ng/mL EGF (Peprotech).

Garancher A., Suzuki H., Haricharan S., Chau L.Q., Masihi M.B., Rusert J.M., Norris P.S., Carrette F., Romero M.M., Morrissy S.A., Skowron P., Cavalli F.M., Farooq H., Ramaswamy V., Jones S.J., Moore R.A., Mungall A.J., Ma Y., Thiessen N., Li Y., Morcavallo A., Qi L., Kogiso M., Du Y., Baxter P., Henderson J.J., Crawford J.R., Levy M.L., Olson J.M., Cho Y.J., Deshpande A.J., Li X.N., Chesler L., Marra M.A., Wajant H., Becher O.J., Bradley L.M., Ware C.F., Taylor M.D, & Wechsler-Reya R.J. (2020). Tumor Necrosis Factor Overcomes Immune Evasion in p53-Mutant Medulloblastoma. Nature neuroscience, 23(7), 842-853.

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Glioblastoma Cell Culture Protocols

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A172, T98G, and U87MG glioblastoma cell lines were purchased from ATCC. Cells were cultured in DMEM media (ThermoScientific) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (P/S, ThermoScientific). GBM4, GBM8, GSC0131, and GSC0827 glioma stem cell (GSC) cultures have been previously described⁵³ (link)^,⁸⁸ (link)^,⁸⁹ (link) and were provided by Drs. Robert Rostomily and Andrei Mikheev, University of Washington and Houston Methodist Hospital (GBM4 and GBM8) and Dr. Patrick Paddison, Fred Hutchinson Cancer Research Center (GSC0131 and GSC0827). GSC cultures were maintained in a defined serum-free medium at 37C and 5% O₂ to mimic in vivo conditions. GBM4 and GBM8 were cultured in Neurobasal medium (ThermoScientific) supplemented with B-27 and N2 (ThermoScientific), 20 ng/mL EGF (PeproTech), 20 ng/mL FGF (PeproTech) and 5 μg/mL heparin (Sigma). GSC0131 and GSC0827 were cultured in Neurocult medium (StemCell Technologies) supplemented with 20 ng/mL EGF (PeproTech), 20 ng/mL FGF (PeproTech), and 0.8 μg/mL heparin (Sigma). All cultures were negative for Mycoplasma contamination.

McFaline-Figueroa J.L., Srivatsan S., Hill A.J., Gasperini M., Jackson D.L., Saunders L., Domcke S., Regalado S.G., Lazarchuck P., Alvarez S., Monnat RJ J.r., Shendure J, & Trapnell C. (2024). Multiplex single-cell chemical genomics reveals the kinase dependence of the response to targeted therapy. Cell Genomics, 4(2), 100487.

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HCC Spheroid Formation Protocol

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A total of 5 × 10³HCC cells were inoculated into six-well ultra-low adsorption plates (Corning, Painted Post, NY, United States) containing 2% B27 (BD PharMingen, Carlsbad, CA, United States), 20 ng/mL epidermal growth factor, 20 ng/mL fibroblast growth factor in DMEM/F12 serum-free medium (Invitrogen). The cells were cultured for 14 d and then imaged and counted under a light microscope.

Chen Q., Yang S.B., Zhang Y.W., Han S.Y., Jia L., Li B., Zhang Y, & Zuo S. (2022). miR-3682-3p directly targets FOXO3 and stimulates tumor stemness in hepatocellular carcinoma via a positive feedback loop involving FOXO3/PI3K/AKT/c-Myc. World Journal of Stem Cells, 14(7), 539-555.

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