Fibroblast growth factor 2 (fgf2)

Manufactured by Abcam

Sourced in United Kingdom, United States, Germany

FGF2 is a recombinant human fibroblast growth factor 2 protein. FGF2 is a member of the fibroblast growth factor family and is involved in various cellular processes, including cell growth, differentiation, and angiogenesis.

Automatically generated - may contain errors

Lab products found in correlation

β mercaptoethanol, by Thermo Fisher Scientific (9 mentions) L glutamine, by Thermo Fisher Scientific (6 mentions) Penicillin streptomycin, by Harvard Bioscience (5 mentions) Vascular endothelial growth factor (vegf), by Abcam (5 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (4 mentions) Gelatin, by Merck Group (3 mentions) Hepatocyte growth factor (hgf), by Abcam (3 mentions) Amphotericin b, by Harvard Bioscience (2 mentions) T75 cell culture flasks, by Sarstedt (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) T75 cell culture flask, by Thermo Fisher Scientific (2 mentions) Ficoll paque plus, by GE Healthcare (2 mentions) Collagenase 4, by Thermo Fisher Scientific (2 mentions) Vegfa, by Abcam (2 mentions) Hif 1α, by Abcam (2 mentions) Image analyzer, by Kodak (2 mentions) Hif 1a, by Novus Biologicals (2 mentions) Peroxidase conjugated anti mouse igg or anti rabbit igg and enhanced chemiluminescence, by Cytiva (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions)

32 protocols using fibroblast growth factor 2 (fgf2)

Directed Differentiation of hPSCs into Pancreatic Progenitors

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hiPSCs and hESCs were differentiated into pancreatic progenitors via modification of previously described methods.^{11 (link)–13 (link)} In brief, hPSCs were treated with Collagenase IV (Life Technologies, Waltham, Massachusetts) and Dispase (STEMCELL Technologies, Vancouver, BC, Canada) at a 1:1 ratio for 3–5 minutes before being manually passaged and passed through a 70 μm cell strainer to remove the mouse embryonic fibroblasts (MEFs). hPSCs were then differentiated 2 days later in RPMI 2% B27 (without vitamin A) (Gibco, Waltham, Massachusetts) supplemented with 100 ng/mL Activin A (R&D Systems, Minneapolis, Minnesota), 3 μM CHIR99021 (Tocris Bioscience, Bristol, UK), 10 μM LY294002 (LC Labs, Woburn, Massachusetts) for 3 days, 50 ng/mL Activin A for 2 days, 50 ng/mL FGF2, 3 μM all-trans retinoic acid (RA) (WAKO, Osaka, Japan), 10 mM nicotinamide (Nic) (Sigma Aldrich, Singapore) for 5 days, 50 ng/mL FGF2, 3 μM RA, 10 mM Nic, 20 μM DAPT (Abcam, Cambridge, UK) for 4 days and 50 ng/mL FGF2, 10 mM Nic, 20 μM DAPT for 3 days to derive pancreatic progenitors.
hPSCs were also differentiated into SC-β cells following the differentiation protocol by Pagliuca et al^{7 (link)} on low attachment plates with shaking for the duration of the whole differentiation protocol, without transfer to a bioreactor.

Loo L.S., Vethe H., Soetedjo A.A., Paulo J.A., Jasmen J., Jackson N., Bjørlykke Y., Valdez I.A., Vaudel M., Barsnes H., Gygi S.P., Ræder H., Teo A.K, & Kulkarni R.N. (2020). Dynamic proteome profiling of human pluripotent stem cell-derived pancreatic progenitors. Stem cells (Dayton, Ohio), 38(4), 542-555.

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Standardized Cultivation of Mesenchymal Stem Cells

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MSCs were harvested and cultivated as published previously, following standardized protocols [58 (link),59 (link)]. Prior to use in the experiments, frozen cells were thawed at 37 °C and resuspended in culture medium. After centrifugation and aspiration of the supernatant medium, cells were expanded in cell culture flasks (Thermo Fisher Scientific, Dreieich, Germany) containing expansion medium (ESM; 83% Dulbecco’s modified Eagle’s medium high glucose, 14% fetal calf serum, 2 mM l-glutamine, 1% non-essential amino acids, 50 µM β-mercaptoethanol (all Thermo Fisher Scientific, Dreieich, Germany), 100 µg/mL penicillin/streptomycin, 2.5 mg/mL amphotericin B (both Biochrom, Berlin, Germany), and 4 ng/mL fibroblast growth factor 2 (Abcam, Berlin, Germany)). Cells in Passage 2 were used for the experiments.

Westhauser F., Essers C., Karadjian M., Reible B., Schmidmaier G., Hagmann S, & Moghaddam A. (2019). Supplementation with 45S5 Bioactive Glass Reduces In Vivo Resorption of the β-Tricalcium-Phosphate-Based Bone Substitute Material Vitoss. International Journal of Molecular Sciences, 20(17), 4253.

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Isolation and Expansion of BMSCs

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The isolation of BMSCs was performed following a density gradient centrifugation protocol as published previously^{33 (link)–35 (link)}. After extracting mononuclear cells from donor bone marrow, cell cultivation was performed in 0.1% gelatin (Sigma Aldrich, Steinheim, Germany) coated T75 cell culture flasks (Sarstedt, Nümbrecht, Germany) in expansion medium (EM), consisting of Dulbecco’s modified Eagle’s medium (DMEM) high glucose, 12.5% fetal calf serum (FCS), 2 mM L-glutamine, 1% non-essential amino acids (NEAA), 50 µM β-mercaptoethanol (all Life Technologies, Darmstadt, Germany), 100 µg/ml penicillin/streptomycin (Sigma-Aldrich) and 4 ng/ml fibroblast growth factor 2 (Abcam, Cambridge, U.K.) under standard cell culture conditions (37 °C and 5% CO₂ in a humidified atmosphere). Medium was exchanged after 24 h to discard non-adherent cells and subsequently twice per week. At 80% confluency, cells were passaged and stored in liquid nitrogen. The experiments were conducted with BMSCs in passage 3.

Decker S., Arango-Ospina M., Rehder F., Moghaddam A., Simon R., Merle C., Renkawitz T., Boccaccini A.R, & Westhauser F. (2022). In vitro and in ovo impact of the ionic dissolution products of boron-doped bioactive silicate glasses on cell viability, osteogenesis and angiogenesis. Scientific Reports, 12, 8510.

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Isolation and Expansion of Bone Marrow-Derived Mesenchymal Stem Cells

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BMSCs were obtained from bone marrow washouts and isolated as described previously [7 (link),23 (link),24 (link)]. In short, after collection in a heparinized syringe, the bone marrow was fractioned on a Ficoll-Paque Plus density gradient (GE Healthcare Europe, Freiburg, Germany). The fraction of mononuclear cells that contains the BMSCs was washed in PBS (Life Technologies) and seeded in T75 cell culture flasks (Nunc, Roskilde, Denmark) coated with 0.1% gelatin (Sigma–Aldrich, Steinheim, Germany). An expansion medium composed of Dulbecco’s Modified Eagle’s Medium (DMEM) high glucose supplemented with 12.5% Fetal Calf Serum (FCS), 1% L-glutamine, 1% non-essential amino acids (NEAA; all Life Technologies), 1% penicillin/streptomycin (Biochrom, Berlin, Germany), 0.1% β-mercaptoethanol (Life Technologies) and 4 ng/mL fibroblast growth factor 2 (Abcam, Cambridge, UK) was used for cell cultivation. Media were changed after the first 24 h to discard non-adherent cells and subsequently twice weekly. Cells were passaged at 80% confluency until usage in passages 4 and 5.

Tsitlakidis S., Hohenbild F., Saur M., Moghaddam A., Kunisch E., Renkawitz T., Gonzalo de Juan I, & Westhauser F. (2023). Reduced Sodium Portions Favor Osteogenic Properties and Cytocompatibility of 45S5-Based Bioactive Glass Particles. Biomimetics, 8(6), 472.

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Isolation and Culture of Bone Marrow Stromal Cells

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The isolation of BMSCs was performed following established protocols [23 (link),24 (link),25 (link)]. In short, mononuclear cells from donor bone marrow were extracted using a density gradient centrifugation protocol and seeded in 0.1% gelatin (Sigma-Aldrich)-coated T75 cell culture flasks (Sarstedt, Nümbrecht, Germany). Cells were cultivated in expansion medium (EM), consisting of DMEM high glucose, 12.5% v/v fetal calf serum (FCS), 2 mM L-glutamine, 1% v/v non-essential amino acids (NEAA), 50 µM β-mercaptoethanol (all Life Technologies, Darmstadt, Germany), 100 µg/mL penicillin/streptomycin (Sigma-Aldrich) and 4 ng/mL fibroblast growth factor 2 (Abcam, Cambridge, UK) at 37 °C and 5% CO₂ in a humidified (≥95%) atmosphere. A first medium exchange was conducted after 24 h to wash off non-adherent cells. Thereafter, medium exchanges were performed twice weekly. Cells were passaged at 80% confluency and transferred to liquid nitrogen storage until further use. The experiments were conducted with BMSCs in passage 4.

Westhauser F., Decker S., Nawaz Q., Rehder F., Wilkesmann S., Moghaddam A., Kunisch E, & Boccaccini A.R. (2021). Impact of Zinc- or Copper-Doped Mesoporous Bioactive Glass Nanoparticles on the Osteogenic Differentiation and Matrix Formation of Mesenchymal Stromal Cells. Materials, 14(8), 1864.

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Isolation and Expansion of Bone Marrow-Derived Mesenchymal Stem Cells

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The BMSCs were harvested from bone marrow and subsequently isolated following well-established protocols [13 (link)]. In short, bone marrow was obtained during surgery using a heparinized syringe. The heparinized bone marrow components were subsequently layered onto Ficoll-Paque Plus (GE Healthcare Europe, Freiburg, Germany) for phase separation. After centrifugation and repeated washing, the mononuclear cells were transferred to T75 culture flasks (Nunc, Roskilde, Denmark), which were coated with 0.1% gelatin (Sigma-Aldrich). Cultivation occurred under standard cell culture conditions (37 °C and 5% CO₂ in a humidified atmosphere) in expansion medium (EM) that consisted of high-glucose DMEM supplemented with 12.5% fetal calf serum (FCS), 2 mM L-glutamine, 1% non-essential amino acids (NEAA), 50 μM β-mercaptoethanol (all Thermo Fisher), 100 μg/mL penicillin/streptomycin (PenStrep; Biochrom, Berlin, Germany), and 4 ng/mL fibroblast growth factor 2 (Abcam, Berlin, Germany). The first exchange of medium was performed after 24 h of cultivation to remove the non-adherent cells. The EM was changed every third day until 80% confluency was observed, and the cells were thereupon passaged. Experiments were conducted with cells in passages 4 and 5.

Rehder F., Arango-Ospina M., Decker S., Saur M., Kunisch E., Moghaddam A., Renkawitz T., Boccaccini A.R, & Westhauser F. (2024). The Addition of Zinc to the ICIE16-Bioactive Glass Composition Enhances Osteogenic Differentiation and Matrix Formation of Human Bone Marrow-Derived Mesenchymal Stromal Cells. Biomimetics, 9(1), 53.

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Isolation and Expansion of Bone Marrow-Derived Mesenchymal Stem Cells

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BMSCs were isolated from bone marrow washouts as described previously [22 (link),23 (link),24 (link)]. In short, bone marrow was collected in a heparinized syringe and cells were fractioned on a Ficoll-Paque Plus density gradient (GE Healthcare Europe, Freiburg, Germany). The fraction of mononuclear cells containing the BMSCs was washed in PBS (Life Technologies, Darmstadt, Germany) and seeded in several 0.1% gelatin-coated (Sigma-Aldrich, Steinheim, Germany) T75 cell culture flasks (Nunc, Roskilde, Denmark).
Cells were cultivated in expansion medium composed of DMEM high glucose supplemented with 12.5% FCS, 1% penicillin/streptomycin, 1% L-glutamine (Life Technologies, Darmstadt, Germany), 1% non-essential amino acids (NEAA; Life Technologies, Darmstadt, Germany), 0.1% β-mercaptoethanol (Life Technologies, Darmstadt, Germany) and 4 ng/mL fibroblast growth factor 2 (Abcam, Cambridge, U.K.). To discard non-adherent cells, medium was changed after the first 24 h and subsequently twice weekly. Cells were passaged at 80% confluency and stored in liquid nitrogen until further use in passage 4.

Hohenbild F., Arango-Ospina M., Moghaddam A., Boccaccini A.R, & Westhauser F. (2020). Preconditioning of Bioactive Glasses before Introduction to Static Cell Culture: What Is Really Necessary?. Methods and Protocols, 3(2), 38.

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Isolation and Expansion of Bone Marrow-Derived MSCs

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Isolation of MSCs from bone marrow was conducted as published previously [[33] , [34] (link), [35] ]. In short, directly after harvesting and washing with 1x phosphate-buffered saline (PBS; Life Technolgies, Darmstadt, Germany), bone marrow was transferred to 0.1% gelatin-coated (Sigma-Aldrich, Steinheim, Germany) culture flasks (Life Technolgies, Darmstadt, Germany) with expansion medium containing 83% Dulbecco's modified Eagle's medium (DMEM) high glucose, 12.5% fetal calf serum (FCS), 2 mM l-glutamine, 1% non-essential amino acids (NEAA), 50 μM β-mercaptoethanol (all Life Technologies, Darmstadt, Germany), 100 μg/ml penicillin/streptomycin, 2.5 mg/ml amphotericin B (both Biochrom, Berlin, Germany) and 4 ng/ml fibroblast growth factor 2 (Abcam, Cambridge, U.K.). After 24 h, the medium was changed in order to remove non-adherent cells and remaining bone marrow. Cultivation was continued with medium changes twice a week until 80% confluence of the cells was reached. The MSCs were transferred to the next passage and repetitive passaging was performed each time the cells reached 80% confluence. MSCs of the donors in passage 1 were pooled in order to reduce inter-individual variances in MSC behavior [35 ]. The pooled cell population was cultivated and cells in passage 4 were used for the experiments.

Schmitz S.I., Widholz B., Essers C., Becker M., Tulyaganov D.U., Moghaddam A., Gonzalo de Juan I, & Westhauser F. (2020). Superior biocompatibility and comparable osteoinductive properties: Sodium-reduced fluoride-containing bioactive glass belonging to the CaO–MgO–SiO2 system as a promising alternative to 45S5 bioactive glass. Bioactive Materials, 5(1), 55-65.

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Isolation and Expansion of Mesenchymal Stem Cells

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MSCs were isolated from freshly obtained bone marrow and prepared as described previously [32 (link),33 (link),34 (link)]. In short, cells were washed in phosphate-buffered saline (PBS; Life Technologies, Darmstadt, Germany) and cultivated in expansion medium consisting of 83% Dulbecco’s modified Eagle’s medium (DMEM), 12.5% fetal calf serum (FCS), 2 mM L-glutamine, 1% non-essential amino acids (NEAA), 50 µM β-mercaptoethanol (all Life Technologies), 100 µg/mL penicillin/streptomycin (Biochrom, Berlin, Germany), and 4 ng/mL fibroblast growth factor 2 (Abcam, Cambridge, United Kingdom). Medium was changed after 24 h and subsequently twice per week. Cells were passaged at 80% confluency and used in passage 5 after being pooled in passage 1 according to previously published recommendations [32 (link)].

Westhauser F., Hohenbild F., Arango-Ospina M., Schmitz S.I., Wilkesmann S., Hupa L., Moghaddam A, & Boccaccini A.R. (2020). Bioactive Glass (BG) ICIE16 Shows Promising Osteogenic Properties Compared to Crystallized 45S5-BG. International Journal of Molecular Sciences, 21(5), 1639.

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Expansion and Cryopreservation of MSCs

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MSCs were extracted from freshly collected bone marrow and cultivated as described previously [74 (link),75 (link),76 (link)]. Cells were washed in phosphate-buffered saline (PBS; Life Technologies, Darmstadt, Germany) and afterwards cultivated in expansion medium composed of 83% DMEM high glucose supplemented with 12.5% FCS, 1% L-glutamine, 1% non-essential amino acids (NEAA; all Life Technologies), 1% penicillin/streptomycin (Biochrom, Berlin, Germany), 0.1% β-mercaptoethanol (Life Technologies) and 4 ng/mL fibroblast growth factor 2 (Abcam, Cambridge, UK). Medium was first changed after 24 h to wash out non-adherent cells. Subsequently medium was changed twice weekly until a confluency of 80% was achieved. The individual donor cells were pooled in passage 1 following previously published recommendations [74 (link)]. Cells were stored in liquid nitrogen until usage in passage 5.

Hohenbild F., Arango Ospina M., Schmitz S.I., Moghaddam A., Boccaccini A.R, & Westhauser F. (2021). An In Vitro Evaluation of the Biological and Osteogenic Properties of Magnesium-Doped Bioactive Glasses for Application in Bone Tissue Engineering. International Journal of Molecular Sciences, 22(23), 12703.

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