Basic fgf

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Israel

Basic FGF is a recombinant protein that acts as a growth factor for various cell types. It is a member of the fibroblast growth factor family and plays a role in cell proliferation, differentiation, and migration. The core function of Basic FGF is to stimulate cell growth and development.

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Recombinant human FGF-basic (41 citations) Human FGF-basic (39 citations) Recombinant human FGF-basic (b-FGF, (6 citations) Recombinant murine FGF-basic (5 citations) Human basic fibroblast growth factor FGF (4 citations) Recombinant Human FGF-basic (154 a.a.) (3 citations) Recombinant Human FGF-basic (100-18B) (3 citations) Human FGF-basic(FGF2) (2 citations) Basic FGF (b-FGF), (2 citations) Human FGF2 (FGF-basic) (154 a.a.) (2 citations)

180 protocols using basic fgf

Directed Differentiation of iPSCs into Hematopoietic Cells

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The cultured iPSCs were treated for up to 16 days (Fig. 2). In summary, on day 1 post-culture, medium was aspirated from each culture and replenished with 500 μl StemDiff APEL medium with BMP-4 (50 ng/ml; PeproTech; Rocky Hill, NJ, USA) and Activin-A (50 ng/ml; PeproTech). On day 3 post-culture, all medium was aspirated from each culture and replenished with 500 μl StemDiff APEL medium with basic-FGF (50 ng/ml; ThermoFisher) and VEGF (50 ng/ml; PeproTech). On day 5 post-culture, all medium was aspirated from each culture and replenished with 500 μl StemDiff APEL medium with basic-FGF, VEG-F, and SB431542 (20 μmol/l; Selleck Chemical LLC). On day 7 post-culture, all medium was aspirated from each culture and replenished with 500 μl StemDiff APEL medium with SCF (50 ng/ml; PeproTech), TPO (50 ng/ml; PeproTech), IL-3 (50 ng/ml; PeproTech), basic-FGF (10 ng/ml; ThermoFisher), VEGF (10 ng/ml; PreproTech), and Flt-3-Ligand (10 ng/ml; PeproTech). Subsequently, every 2--3 days, medium was aspirated from each culture and replenished with the complete medium described for day 7. From days 11-16, nonadherent CD34 + cells, released from the attached cells, were collected for flow cytometry analysis and semi-solid MethoCult culture.

Teque F., Ye L., Xie F., Wang J., Morvan M.G., Kan Y.W, & Levy J.A. (2020). Genetically-edited induced pluripotent stem cells derived from HIV-1-infected patients on therapy can give rise to immune cells resistant to HIV-1 infection. AIDS (London, England), 34(8).

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Derivation and Maintenance of Wnt3-null Epiblast Stem Cells

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E6.0 embryos were isolated in M2 medium; extraembryonic tissue and primitive endoderm were removed. Explants were cultured on mitotically inactivated mouse embryonic fibroblasts (MEFs) in medium containing 20% KOSR, 77% DMEM/F12, 100 U/mL 1× penicillin/streptomycin (pen/strep), 2 mM L-glutamine, 1× NEAAs, 0.01% β-mercaptoethanol (β-ME), 12 ng/mL Fgf basic (ThermoFisher), 20 ng/mL Activin A (R&D Systems), and 2 µM Wnt inhibitor IWP2 (Sigma Aldrich) in DMSO. Wnt3-null EpiSCs were genotyped once cultured free of MEFs. MEF-free culture took place on a surface coated by fibronectin (ThermoFisher) in medium containing 48% DMEM/F12, 48% neurobasal medium, 1% B27 supplement, 0.5% N2 supplement B (all ThermoFisher), 1× pen/strep, 12 ng/mL Fgf basic, 20 ng/mL Activin A, 2 µM IWP2, and 0.01% β-ME. Medium was refreshed on a daily basis. Cells were tested for mycoplasma contamination. Wnt stimulation was performed after at least two passages of feeder-free culturing by withdrawal of IWP2 and addition of 3 µM Chiron.

Neijts R., Amin S., van Rooijen C., Tan S., Creyghton M.P., de Laat W, & Deschamps J. (2016). Polarized regulatory landscape and Wnt responsiveness underlie Hox activation in embryos. Genes & Development, 30(17), 1937-1942.

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Stepwise Human ESC Differentiation to Isthmic Organizer-like Cells

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Human ESCs (H9, WiCell Inc., USA) were cultured in human ESC medium composed of DMEM/F12 medium (Invitrogen, USA) supplemented with 20% Knockout-Serum Replacement (Invitrogen), 1% nonessential amino acids (Invitrogen), 0.1 mM beta-mercaptoethanol (Sigma, USA), and 4 ng/mL basic FGF (Peprotech, USA). For differentiation, embryoid bodies (EBs) were formed by mechanically detaching ESC colonies and culturing them in DMEM/F12:Neurobasal media (Invitrogen) (1:1), 1% N2 supplement (Invitrogen), and 2% B27 supplement without vitamin A (Invitrogen). On day 4 of differentiation, these EBs were plated onto Matrigel (BD Biosciences, USA)-coated dishes and cultured in the same medium except that the concentrations of N2 and B27 supplements were reduced by half (0.5%) for five days. During the first four days, 5 μM dorsomorphin (Calbiochem, USA) and 10 μM SB431542 (SB) (Sigma) were added to the medium to facilitate neural induction. To induce IsO-like cells, CHIR99021 (CHIR) at various concentrations (0–1.2 μM) (Calbiochem) and 100 ng/ml FGF8 (Peprotech) were added to the medium as described in Supplementary Fig. S1.

Lee J., Choi S.H., Lee D.R., Kim D.S, & Kim D.W. (2018). Generation of Isthmic Organizer-Like Cells from Human Embryonic Stem Cells. Molecules and Cells, 41(2), 110-118.

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Oncosphere Culture Assay

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These assays were performed as described previously [33 (link), 34 (link)]. Cells were seeded on ultra-low attachment culture dishes (Corning, Corning, NY) in serum-free DMEM-F12 medium containing 50 μg/ml insulin (Sigma-Aldrich St. Louis, MO), 0.4% Albumin Bovine Fraction V (Sigma-Aldrich St. Louis, MO), N^− 2 Plus Media Supplement (Life Technologies, Grand Island, NY), B-27 Supplement (Life Technologies, Grand Island, NY), 20 μg/ml EGF (PeproTech Rocky Hill, NJ), and 10 μg/ml basic FGF (PeproTech, Rocky Hill, NJ) to support the growth of undifferentiated oncospheres. Cells were incubated in a CO2 incubator for 1–2 weeks, and the numbers of oncosphere cells were counted under a microscope.

Yang R., Liu N., Chen L., Jiang Y., Shi Y., Mao C., Liu Y., Wang M., Lai W., Tang H., Gao M., Xiao D., Wang X., Yu F., Cao Y., Yan Q., Liu S, & Tao Y. (2019). LSH interacts with and stabilizes GINS4 transcript that promotes tumourigenesis in non-small cell lung cancer. Journal of Experimental & Clinical Cancer Research : CR, 38, 280.

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Establishing Glioma Neurosphere Cultures

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Brains of tumor-bearing mice were micro-dissected and glioma tissue was mechanically and enzymatically disaggregated for the generation of ex-vivo cultures. The cultures were plated as single cells in serum-free DMEM/F-12 1:1 media (Life Technologies) supplemented with B27 without Vitamin A (Life Technologies), 10 ng/mL EGF (Peprotech), and 10 ng/mL basic FGF (Peprotech), and grown as tumor neurospheres. Every 3–4 days, the cells were dissociated by triturating with Accutase (Sigma-Aldrich) and sub-cultured, as described previously [39 (link)]. All cell lines were routinely mycoplasma tested using the MycoAlert mycoplasma detection kit (Lonza). For serum-induced differentiation studies, the neurosphere cultures were dissociated into single cells and the growth medium was switched to Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS (Hyclone). The cells were grown under these conditions for 7–10 days and assessed for expression of differentiation markers. For sphere formation assays, cells were dissociated and plated at single-cell density in Nunclon Sphera Surface ultra-low adhesion 96-well plates (Thermo Fisher) in neurosphere medium. Sphere numbers per 96-well plate were quantified after 12 days. Primary astrocytes were isolated from 3-day old pups and cultured in DMEM supplemented with 10% FBS.

Todorova P.K., Fletcher-Sananikone E., Mukherjee B., Kollipara R., Vemireddy V., Xie X.J., Guida P.M., Story M.D., Hatanpaa K., Habib A.A., Kittler R., Bachoo R., Hromas R., Floyd J.R, & Burma S. (2019). Radiation-induced DNA damage cooperates with heterozygosity of TP53 and PTEN to generate high grade gliomas. Cancer research, 79(14), 3749-3761.

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Tumorsphere Formation Assay for LNCaP-abl Cells

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LNCaP-abl cells were digested, and 2000 cells were seeded into ultralow attachment 6-well plates (Corning Inc., Life Sciences, USA) with DMEM/F-12 (HyClone) serum-free medium containing 1 μL/mL transferrin (Sigma-Aldrich, USA), 20 ng/mL EGF (PeproTech, USA), 20 ng/mL basic FGF (PeproTech), 2% B27 (Invitrogen, USA), and 10 unit/mL human LIF (Sigma-Aldrich). The formed tumorspheres were counted by inverse microscopy after 2 weeks.

Lin Q., Cao J., Du X., Yang K., Yang X., Liang Z., Shi J, & Zhang J. (2022). CYP1B1-catalyzed 4-OHE2 promotes the castration resistance of prostate cancer stem cells by estrogen receptor α-mediated IL6 activation. Cell Communication and Signaling : CCS, 20, 31.

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Isolation and Culture of Mouse Cortical Neural Precursor Cells

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Dorsal forebrains from timed-pregnant E13.5 mouse embryos were digested with Accutase (Fisher) to yield dissociated cortical neural precursor cells (NPCs) for culture. NPCs were carried on plates coated with Matrigel (Corning) at 80 μg/ml and maintained in DMEM-F12 medium (GIBCO) supplemented with B27 (GIBCO), N2 (GIBCO), and Glutamax (GIBCO). A growth factor cocktail containing EGF (PeproTech) (20ng/ml) and basic FGF (PeproTech) (20ng/ml) in Heparin (5 μg/ml) was added to the medium fresh. Cells were carried at densities not exceeding 80%, and all experiments were performed on density- and passage-matched NPC cultures. Cells were incubated in standard conditions: 37°C with 5% CO₂. Multiple lines of NPCs were generated from independent litters of Fmr1 wild-type and knockout embryos.

Edens B.M., Vissers C., Su J., Arumugam S., Xu Z., Shi H., Miller N., Rojas Ringeling F., Ming G.L., He C., Song H, & Ma Y.C. (2019). FMRP Modulates Neural Differentiation through m6A-Dependent mRNA Nuclear Export. Cell reports, 28(4), 845-854.e5.

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3D Spheroid Culture and Labeling

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MCF-7 and A549 spheroids were obtained by plating cells at low density (1000 cells/cm²) on poly-HEMA coated wells. Spheroids were growth in DMEM/Neurobasal medium (1:1) added with Glutamax (Gibco, Thermo Fisher Scientific, Waltham, MA USA), 100 μg/mL penicillin/streptomycin, 1% B-27 and N2 supplements (Gibco, Thermo Fisher Scientific, Waltham, MA USA), 10 ng/mL basic FGF and 20 ng/mL EGF (Peprotech EC, London, UK). The spheroids were maintained in culture for 21 days before the experiments and passaged at 70% confluence using Accutase dissociation reagent (Sigma-Merck, Darmstadt, Germany) and split at a 1:3 ratio. Before the experiments, the spheroids were gently dissociated, the cells were loaded by 5 μM calcein-AM (BD Italy, Milan, Italy for 30 min and then injected in the circuit.

Paolillo M., Colombo R., Serra M., Belvisi L., Papetti A., Ciusani E., Comincini S, & Schinelli S. (2019). Stem-Like Cancer Cells in a Dynamic 3D Culture System: A Model to Study Metastatic Cell Adhesion and Anti-Cancer Drugs. Cells, 8(11), 1434.

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Isolation and Expansion of Nucleus Pulposus Cells

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NP cells were isolated using a modified method as described before [26 (link)]. In brief, the tissue was digested with collagenase CLS II (333.3 U/mL) (Biochrom), collagenase P (1 U/mL) (Roche, Mannheim, Germany), and hyaluronidase (33.3 U; Roche) for 2–4 h under continuous stirring in a spinner flask at 37°C and 5% CO₂. After digestion, the cells were plated with a density of 10⁴ cells/cm². The cells were cultivated at 37°C and 5% CO₂ with medium supplemented either with or without 2 ng/mL basicFGF (Peprotech, Hamburg, Germany). The medium was changed every 2–3 days. When reaching about 80–90% confluence the cells were passaged using trypsin/ethylenediaminetetraacetic acid (EDTA; Biochrom). For subsequent passages, the seeding density was 5.000 cells/cm². For the tissue engineered grafts, cells were used at the end of passage 2. For investigation of growth kinetics, the NP cells were cultivated up to passage 7 with or without 2 ng/mL basicFGF supplementation with a seeding density of 8.000 cells/cm² at passage 1 and following passages. The volume of medium was 25 mL in each cell culture flask with a surface of 175 cm².

Stich S., Stolk M., Girod P.P., Thomé C., Sittinger M., Ringe J., Seifert M, & Hegewald A.A. (2015). Regenerative and Immunogenic Characteristics of Cultured Nucleus Pulposus Cells from Human Cervical Intervertebral Discs. PLoS ONE, 10(5), e0126954.

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Culturing Hippocampal Neurospheres from WT and Cln5-KO Mice

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The hippocampi were dissected from the brains of WT and Cln5-KO mice at E18 and cultured as described (Kärkkäinen et al., 2014 (link)) as free-floating neurospheres in the presence of epidermal growth factor (EGF) and basic fibroblastic growth factor (FGF). Cells were grown in culture medium containing DMEM/F12 (Gibco, Thermo Fisher Scientific), 1 M HEPES (Sigma-Aldrich), 100 U/ml penicillin, 100 mg/ml streptomycin, B27 supplement (all from Gibco, Thermo Fisher Scientific), 20 ng/ml EGF (PeproTech, Rocky Hill, NJ, USA) and 10 ng/ml basic FGF (PeproTech). Cells were cultivated at 37°C in 5% CO₂. Fresh medium as well as EGF and FGF were added to the cells every 3 days.

Savchenko E., Singh Y., Konttinen H., Lejavova K., Mediavilla Santos L., Grubman A., Kärkkäinen V., Keksa-Goldsteine V., Naumenko N., Tavi P., White A.R., Malm T., Koistinaho J, & Kanninen K.M. (2017). Loss of Cln5 causes altered neurogenesis in a mouse model of a childhood neurodegenerative disorder. Disease Models & Mechanisms, 10(9), 1089-1100.

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