NIH 3T3 mouse fibroblast cell lines (ATCC CRL-1658) were cultured in 100 × 20 mm and 150 × 25 mm culture dishes (Falcon Corning cat. no. 353003 and 353025) using Dulbecco’s modified Eagle’s medium containing 4.5 g/L D-glucose, L-glutamine, 110 mg/L sodium pyruvate (DMEM, GIBCO 11995-065) with 10% heat inactivated newborn calf serum, NBCS (GIBCO 26010-074). Primary pancreatic stellate cells (PSC) were cultured until passage 8 in DMEM with 10% heat inactivated fetal bovine serum and penicillin-streptomycin. Cultures were allowed to reach 80% confluency before passaging them into new media. Briefly, cells were washed twice with phosphate-buffered saline (PBS) without calcium and magnesium, Wisent 311-010-CL, cells detached using Trypsin-0.25% EDTA (Wisent) and split at a seeding density of 5,000 cells/cm2. Commercial GFs used in this study included the following: human FGF1 (PeproTech, cat. no. 100–17A), human FGF2 (PeproTech, cat. no. 100–18B), human PDGF-BB (PeproTech, cat. no. 100–14B), human IGF1 (PeproTech, cat. no. 100-11), human IGF2 (PeproTech, cat. no. 100-12), human TGF-β1(PeproTech, cat. no. 100–21C).
Human fgf1
Manufactured by Thermo Fisher Scientific
Human FGF1 is a recombinant human fibroblast growth factor 1 protein. It functions as a signaling molecule involved in various cellular processes, including cell growth, differentiation, and angiogenesis.
Automatically generated - may contain errors
Lab products found in correlation
Human pdgf bb, by Thermo Fisher Scientific (1 mentions)
Human tgf β1, by Thermo Fisher Scientific (1 mentions)
Human fgf2, by Thermo Fisher Scientific (1 mentions)
Human igf2, by Thermo Fisher Scientific (1 mentions)
Human igf1, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Culture dishes, by Corning (1 mentions)
Optical bottom, by Thermo Fisher Scientific (1 mentions)
Varioskan lux plate reader, by Thermo Fisher Scientific (1 mentions)
Heparin sodium salt, by Merck Group (1 mentions)
Celltiter glo assay, by Promega (1 mentions)
2 protocols using human fgf1
1
Culturing Mouse Fibroblasts and Pancreatic Stellate Cells
2
FGF1-Recifercept Proliferation Assay
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5000 cells were plated in white 96-well plate with optical bottom (Nunc). Prior to plating, cells were centrifuged twice at 800g, 5 minutes at room temperature and washed with assay medium (RPMI-1640, 10% heat inactivated fetal bovine serum, 1% penicillin/streptomycin/glutamine) to remove of IL-3 and puromycin.
Cells were either stimulated for 72h with increasing concentrations of human FGF1 (Peprotech) or simultaneously with 10ng/mL human FGF1 and increasing concentrations of Recifercept, both supplemented with 10μg/mL heparin sodium salt (Sigma) acting as a co-factor. Cell proliferation was assessed by measuring intracellular ATP concentration with CellTiter Glo assay (Promega). Luminescence readout was measured with a Varioskan Lux plate reader (Thermo Fisher Scientific).
Cells were either stimulated for 72h with increasing concentrations of human FGF1 (Peprotech) or simultaneously with 10ng/mL human FGF1 and increasing concentrations of Recifercept, both supplemented with 10μg/mL heparin sodium salt (Sigma) acting as a co-factor. Cell proliferation was assessed by measuring intracellular ATP concentration with CellTiter Glo assay (Promega). Luminescence readout was measured with a Varioskan Lux plate reader (Thermo Fisher Scientific).
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