β fgf

Manufactured by STEMCELL

β‐FGF is a recombinant human fibroblast growth factor-2 (FGF-2) protein. FGF-2 is a key regulator of cell growth, differentiation, and survival. It plays a role in the development and maintenance of various cell types, including endothelial, neural, and stem cells.

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Lab products found in correlation

Epidermal growth factor (egf), by STEMCELL (3 mentions) Dmem f12 medium, by Thermo Fisher Scientific (2 mentions) Ultra low attachment dishes, by Corning (2 mentions) Ampicillin, by Sangon (2 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Mem non essential amino acid, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Knockout serum replacement (ksr), by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Sendai virus reprogramming, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Merck Group (1 mentions) Facsaria 2 cytometer, by BD (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) 40 μm cell strainer, by BD (1 mentions) Trypsin inhibitor, by Worthington (1 mentions) Ampicillin, by Merck Group (1 mentions) Chir99021, by Merck Group (1 mentions) Valproic acid, by Merck Group (1 mentions) Ly411575, by Merck Group (1 mentions)

4 protocols using β fgf

Cochlear Progenitor Cell Culture Protocol

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Cochlear progenitors were cultured in suspension with Dulbecco's Modified Eagle Medium/Nutrient Misxture F‐12 (DMEM/F‐12) medium (ThermoFisher, #11330–032) plus 1% N2 (ThermoFisher, #17502048), 2% B27 (ThermoFisher, #17504044), epidermal growth factor (EGF, 20 ng/mL; StemCell, #78006.1), insulin‐like growth factor (IGF, 50 ng/mL, Sigma, #I8779), basic fibroblast growth factor (β‐FGF, 10 ng/mL, StemCell, #78003) and 0.1% ampicillin (Sangon Biotech, #A610028‐0025) and cultured in ultra‐low attachment dishes (Corning, #3474) for 5 days.
For the differentiation assay, the suspended spheres were transferred to laminin‐coated four‐well dishes and cultured in DMEM/F12 medium (ThermoFisher, #11330–032) with 1% N2 (ThermoFisher, #17502048), 2% B27 (ThermoFisher, #17504044), EGF (20 ng/mL; StemCell, #78006.1), IGF (50 ng/mL, Sigma, #I8779), β‐FGF (10 ng/mL, StemCell, #78003), LY411575 (5 μM; Sigma‐Aldrich, #SML0506), CHIR99021 (3 μM; Sigma‐Aldrich, #SML1046) and 0.1% ampicillin (Sangon Biotech, #A610028‐0025) for 7 days.

Xu C., Qi J., Hu X., Zhang L., Sun Q., Li N., Chen X., Guo F., Wu P., Shi Y, & Chai R. (2023). Rps14 upregulation promotes inner ear progenitor proliferation and hair cell regeneration in the neonatal mouse cochlea. Cell Proliferation, 56(5), e13458.

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Sendai Virus Reprogramming of Fibroblasts

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Fibroblasts were reprogrammed using the Sendai virus reprogramming method as recommended by the manufacturer (Life Technologies). Briefly, fibroblasts were infected using SeV vectors encoding OCT3/4, SOX2, KLF4, and c-MYC on day 0. Two days later, cells were trypsinized and picked up onto two 10-cm gelatin-coated culture dishes that had been seeded with irradiated mouse embryonic fibroblasts (irrMEFs, GlobalStem). The cultures were maintained in human embryonic stem cell (hESC) medium containing DMEM/F12 (Gibco), 20% KnockOut Serum Replacement (Gibco), 1% minimum essential medium (MEM) nonessential amino acid (Gibco), 1 mM l-glutamine (Gibco), 0.1 mM β-mercaptoethanol (Sigma-Aldrich), and 10 ng/mL of basic fibroblast growth factor (β-FGF, Stem Cell technologies). Clones were picked starting on 20 days post infection, and expanded on irrMEFs before being adapted to feeder-free conditions.

Roux G.L., Jarray R., Guyot A.C., Pavoni S., Costa N., Théodoro F., Nassor F., Pruvost A., Tournier N., Kiyan Y., Langer O., Yates F., Deslys J.P, & Mabondzo A. (2019). Proof-of-Concept Study of Drug Brain Permeability Between in Vivo Human Brain and an in Vitro iPSCs-Human Blood-Brain Barrier Model. Scientific Reports, 9, 16310.

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Lgr5-EGFP+ Cochlear Cell Isolation

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The cochleae from P0‐P1 Lgr5‐EGFP‐Cre^ERT2 mice were separated rapidly in pre‐chilled PBS (pH 7.2) and digested with 0.125% trypsin (ThermoFisher, #25200056) plus 1% DNase I (Sigma, #DN25) at 37°C for 8 min. Digestion was terminated with trypsin inhibitor (10 mg/mL, Worthington Biochem, #LS003570), and tissues were mechanically triturated for about 80–100 strokes with blunt pipette tips (Eppendorf, #22491245) into single‐cell suspensions. Isolated cells were percolated through a 40‐μm cell strainer (BD Biosciences, #21008‐949) and then sorted using the GFP channel on a BD FACSAria* II cytometer (BD Biosciences). The flow‐sorted Lgr5‐EGFP⁺ cells were diluted to 500 cells/well and cultured in ultra‐low attachment dishes (Corning, #3474) for 5 days in DMEM/F12 medium (ThermoFisher, #11330‐032) plus 2% B27 (ThermoFisher, #17504044), 1% N2 (ThermoFisher, #17502048), EGF (20 ng/mL; Stem Cell Technologies, #78006.1), IGF (50 ng/mL, Sigma, #I8779), β‐FGF (10 ng/mL, Stem Cell Technologies, #78003) and 0.1% ampicillin (Sangon Biotech, #A610028‐0025).

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Expansion and Differentiation of Cochlear Cells

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The basilar membranes of the cochleae from P2 mice were dissected out and digested into single cells in the same way as for flow cytometry. Cell numbers were counted to establish the total number. Equal numbers of cells were re‐suspended in a 7:3 mixed matrix (Corning, #356231) and expansion medium and plated onto a prewarmed 24 well plate. (Greiner, #662160). The expansion medium was prepared with DMEM/F12 medium (ThermoFisher, #11330–032) with 1% N2 (ThermoFisher, #17502048), 2% B27 (ThermoFisher, #17504044), EGF (20 ng/mL; StemCell, #78006.1), IGF (50 ng/mL, Sigma, #I8779), β‐FGF (10 ng/mL, StemCell, #78003), 0.1% ampicillin (Sangon Biotech, #A610028‐0025), valproic acid (1 mM; Sigma‐Aldrich, #P4543), CHIR99021 (3 μM; Sigma‐Aldrich, #SML1046) and 616452 (2 μM; Sigma‐Aldrich, #446859‐33‐2). After 10 days of expansion, a single 5‐ethynyl‐2′‐deoxyuridine (EdU) pulse (10 mM, ThermoFisher, #C10340) was introduced and then was analysed 1 h later. To induce differentiation, the expansion medium was replaced at Day 10 by differentiation medium composed of DMEM/F12 (ThermoFisher, #11330‐032) with 1% N2 (ThermoFisher, #17502048), 2% B27 (ThermoFisher, #17504044), LY411575 (5 μM; Sigma‐Aldrich, #SML0506), CHIR99021 (3 μM; Sigma‐Aldrich, #SML1046) and 0.1% ampicillin. The culture medium was changed every other day.

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