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Recombinant human fibroblast growth factor 2

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Recombinant human fibroblast growth factor-2 is a protein that promotes cell growth and differentiation. It is a member of the fibroblast growth factor family and is involved in various cellular processes, including angiogenesis, wound healing, and embryonic development.

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Lab products found in correlation

B27 supplement, by Thermo Fisher Scientific (2 mentions) N2 supplement, by Thermo Fisher Scientific (2 mentions) Recombinant human epidermal growth factor, by R&D Systems (2 mentions) Neurobasal medium, by Thermo Fisher Scientific (2 mentions) L glutamine, by Corning (2 mentions) Heparin, by Merck Group (2 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions) High glucose dmem, by Thermo Fisher Scientific (1 mentions) Dmem low glucose, by Merck Group (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Proline, by Merck Group (1 mentions) Ascorbate 2 phosphate, by Merck Group (1 mentions) β glycerol phosphate, by Merck Group (1 mentions) Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions) Low glucose dmem, by Thermo Fisher Scientific (1 mentions) Lookout mycoplasma kit, by Merck Group (1 mentions) Neurocult chemical dissociation kit, by STEMCELL (1 mentions) Penicillin g streptomycin sulfate amphotericin b complex, by Corning (1 mentions) Ec growth supplement, by Thermo Fisher Scientific (1 mentions) Stemspan, by STEMCELL (1 mentions)

Close spelling variants of this product

Human fibroblast growth factor 2 (5 citations) Fibroblast growth factor (FGF)-2 (5 citations) Recombinant human fibroblast growth factor basic (rhFGF-2; (2 citations)

9 protocols using recombinant human fibroblast growth factor 2

1

Isolation and Culture of Human GBM Cells

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Human patient derived high grade glioma cells were isolated from dissociated surgical tumor specimens with approval of Kobe University Hospital Institutional Review Board [37 (link)]. Human patient derived cells were cultured in EF20 medium composed of Neurobasal medium (ThermoFisher Gibco) supplemented with 3 mM l-Glutamine (Corning), 1 × B27 supplement (ThermoFisher Gibco), 0.5 × N2 supplement (ThermoFisher Gibco), 20 ng/ml recombinant human epidermal growth factor (R&D Systems), 20 ng/ml recombinant human fibroblast growth factor-2 (PeproTech), and 0.5 × penicillin G/streptomycin/amphotericin B complex (Corning) at 37 °C and 5% CO2.
Tanaka K., Sasayama T., Nagashima H., Irino Y., Takahashi M., Izumi Y., Uno T., Satoh N., Kitta A., Kyotani K., Fujita Y., Hashiguchi M., Nakai T., Kohta M., Uozumi Y., Shinohara M., Hosoda K., Bamba T, & Kohmura E. (2021). Glioma cells require one-carbon metabolism to survive glutamine starvation. Acta Neuropathologica Communications, 9, 16.
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2

Multilineage Differentiation Protocols for Stem Cells

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Control Culture Medium (CCM) contained low glucose DMEM (Invitrogen, Thermo-Fisher Scientific, Waltham, MA) with 10% lot-selected fetal bovine serum (FBS; Atlanta Biologicals, Flowery Branch, GA), 10 ng/mL recombinant human fibroblast growth factor-2 (Peprotech, Rocky Hill, NJ) and 1% penicillin-streptomycin (P/S) (Invitrogen).
Chondrogenic differentiation medium (Chondro) consisted of high glucose DMEM (Invitrogen) containing 10 ng/mL recombinant human transforming growth factor beta 3 (rhTGF-β3; R&D Systems, Minneapolis, MN), 1% Insulin-Transferrin-Selenium (ITS+ Premix™; BD, San Jose, CA), 100 nM dexamethasone, 50 mg/L ascorbate-2 phosphate, 0.4 mM proline (Sigma-Aldrich, St Louis, MO) and 1% P/S (Invitrogen) (Valonen et al., 2010 (link)).
Osteogenic differentiation medium (Osteo) consisted of low glucose DMEM containing 10% FBS, 1% P/S, 100 nM dexamethasone, 50 μM ascorbate-2 phosphate, and 10 mM beta glycerol phosphate (Sigma-Aldrich), which has been shown to be required for osteogenic differentiation of MSCs (Abrahamsson et al., 2010 (link)).
Larson B.L., Yu S.N., Park H., Estes B.T., Moutos F.T., Bloomquist C.J., Wu P.B., Welter J.F., Langer R., Guilak F, & Freed L.E. (2019). Chondrogenic, hypertrophic, and osteochondral differentiation of human mesenchymal stem cells on three-dimensionally woven scaffolds. Journal of tissue engineering and regenerative medicine, 13(8), 1453-1465.
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3

GBM Stem-like Cells Culture Protocol

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Mouse 005 GBM stem-like cells (GFP positive) were provided by Dr. I Verma (Salk Institute) and have been described [21 (link), 24 (link)]. They were cultured as spheres in EF20 medium composed of Neurobasal medium (Thermo Fisher Gibco) supplemented with 3 mM l-Glutamine (Corning Mediatech), 1 × B27 supplement (Thermo Flasher Gibco), 0.5 × N2 supplement (Thermo Fisher Gibco), 2 μg/ml heparin (Sigma, St Louis, MO), 20 ng/ml recombinant human epidermal growth factor (R&D Systems, Minneapolis, MN), 20 ng/ml recombinant human fibroblast growth factor-2 (PeproTech, Rocky Hill, NJ), and 0.5 × penicillin G/streptomycin sulfate/amphotericin B complex (Corning Mediatech) at 37 °C and 5% CO2. To passage cells, neurospheres were dissociated with the Neurocult chemical dissociation kit (Stem Cell Technologies). Mouse GBM GL261 cells were obtained from the National Cancer Institute and grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal calf serum (FCS). Cells were confirmed to be mycoplasma free (LookOut Mycoplasma kit, Sigma) and used at low passage number.
Li M., Li G., Kiyokawa J., Tirmizi Z., Richardson L.G., Ning J., Das S., Martuza R.L., Stemmer-Rachamimov A., Rabkin S.D, & Wakimoto H. (2020). Characterization and oncolytic virus targeting of FAP-expressing tumor-associated pericytes in glioblastoma. Acta Neuropathologica Communications, 8, 221.
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4

Neuronal Cell Culture Reagents

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The following materials were purchased: Hank’s balanced salt solution, neurobasal medium, B27 and 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) from Invitrogen (Carlsbad, CA, USA); recombinant human fibroblast growth factor-2 from Pepro Tech (London, UK); atelocollagen from Koken (Tokyo, Japan); CORT and sertraline HCl from Tokyo Chemical Industry (Tokyo, Japan).
Kigawa Y., Hashimoto E., Ukai W., Ishii T., Furuse K., Tsujino H., Shirasaka T, & Saito T. (2014). Stem cell therapy: a new approach to the treatment of refractory depression. Journal of Neural Transmission, 121(10), 1221-1232.
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5

Expanding and Differentiating Hematopoietic Stem Cells

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E-SLAM HSPCs were sorted and cultured in StemSpan serum-free expansion medium (SFEM) (Stem Cell Technologies, Vancouver, BC, Canada) containing recombinant mouse SCF (300 ng/ml) and recombinant mouse IL-11 (20 ng/ml) (Stem Cell Technologies). CD41+ MK cells were cultured in SFEM with 25ng/ml recombinant mouse SCF and 25 ng/ml recombinant human thrombopoietin (TPO) (Stem Cell Technologies). MK-conditioned media (MKCM) was collected from MK cells after 48–72 h of culture. ECs were cultivated in advanced DMEM/F12 medium supplemented with 20% fetal bovine serum, 50 µg/ml EC growth supplement (Alfa Aesar, Ward Hill, MA, USA), 10 ng/ml recombinant mouse vascular endothelial growth factor and 20 ng/ml recombinant human fibroblast growth factor 2 (FGF2) (both from PeproTech, Rocky Hill, NJ, USA). Details are provided in Supplementary Information.
Zhan H., Ma Y., Lin C, & Kaushansky K. (2016). JAK2V617F-mutant megakaryocytes contribute to hematopoietic stem/progenitor cell expansion in a model of murine myeloproliferation. Leukemia, 30(12), 2332-2341.
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6

Biomaterial Synthesis and Characterization

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Chitosan (low molecular weight ∼50 kDa), heparin sodium salt from bovine intestinal mucosa, MTT (3-[4,5-dimetylthiazol-2-yl]-2,5-dipheniltetrazolium), 4′, 6′-diamidino-2-phenylindole, and rhodamine-conjugated phalloidin were purchased from Sigma (St Louis, MO). Genipin was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Recombinant human-fibroblast growth factor-2 (FGF-2) was purchased from Peprotech (Rocky Hill, NJ). NT-3 and brain-derived neurotrophic factor (BDNF) were purchased from Alomone Labs (Jerusalem, Israel). Nitrocellulose and NuPAGE 4% to 12% Bis-Tris Gels were purchased from Life Technologies (Carlsbad, CA).
Skop N.B., Singh S., Antikainen H., Saqcena C., Calderon F., Rothbard D.E., Cho C.H., Gandhi C.D., Levison S.W, & Dobrowolski R. (2019). Subacute Transplantation of Native and Genetically Engineered Neural Progenitors Seeded on Microsphere Scaffolds Promote Repair and Functional Recovery After Traumatic Brain Injury. ASN NEURO, 11, 1759091419830186.
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7

Polymer-Mediated Transfection of Stem Cell Regulators

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Homo sapiens SRY (sex determining region Y)-box 2 (Sox2, CMV Promoter, RG200757) and oligodendrocyte transcription factor 2 (Olig2, CMV Promoter, RG208209) as transfection-ready DNAs were obtained from OriGene Technologies (Rockville, MD). pEGFP-N1 DNA was obtained from Elim Biopharmaceuticals (Hayward, CA). Recombinant human fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) were purchased from Peprotech (Rocky Hill, NJ). Poly-l-lysine was obtained from ScienCell Research Laboratories (Carlsbad, CA). 4′,6-Diamidino-2-phenylindole, dihydrochloride (DAPI) was purchased from Molecular Probes (Eugene, OR). Primers and primary antibodies used in this study are shown in Tables S1 and S2,† respectively. Fluorophore-conjugated secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA).
Monomers used for synthesizing polymers (Scheme 1) were obtained as follows: 1,4-butanediol diacrylate (B4; Alfa Aesar, Ward Hill, MA), 1,5-pentanediol diacrylate (B5; Monomer-Polymer and Dajac Labs, Trevose, PA), 3-amino-1-propanol (S3; Alfa Aesar), 4-amino-1-butanol (S4; Alfa Aesar), 5-amino-1-pentanol (S5; Alfa Aesar), 2-(3-aminopropylamino) ethanol (E6; Sigma Aldrich, St Louis, MO), and 1-(3-aminopropyl)-4-methylpiperazine (E7; Alfa Aesar). All other chemical reagents were obtained from Sigma Aldrich (St. Louis, MO).
Li X., Kozielski K., Cheng Y.H., Liu H., Zamboni C.G., Green J, & Mao H.Q. (2016). Nanoparticle-mediated conversion of primary human astrocytes into neurons and oligodendrocytes. Biomaterials science, 4(7), 1100-1112.
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8

Human Glioblastoma Stem Cell Culturing

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Human glioma initiating cells (hGIC) L0306, kindly provided by Dr. R. Galli 16 (link), were engineered with pCAG:DsRed and oCAG:mGFP-Gluc and maintained as spheres in Neurocult Medium supplemented with NeuroCult NS-A (StemCell Technologies, Vancouver, British Columbia, CA), human recombinant fibroblast growth factor 2 (10 ng/ml; PeproTech, Rocky Hill, USA), epidermal growth factor (20 ng/ml; PeproTech, Rocky Hill, USA) and Heparin (2µg/ml, Sigma-Aldrich, Milano,Italy).
HGG[vIII] cells were obtained from INK4a/ARF knock-out BALB/c mice intracranially injected with syngeneic neural progenitor cells transduced with pCAG:DsRed-EGFRvIII as previously described 17 (link). Cells were maintained in Dulbecco's modified Eagle's medium-F12 (Invitrogen, Carlsbad,CA) with B27 supplement (Invitrogen, Carlsbad,CA), human basic fibroblast growth factor (10 ng/ml, Peprotech, London, UK) and epidermal growth factor (10 ng/ml, Peprotech, London, UK) and plated on Matrigel (1:200; BD Biosciences, Franklin Lakes, NJ). Cultures from tumors were established microdissecting DsRed-positive areas under a fluorescence microscope and trypsinizing them for 20 minutes. Cells were maintained in the medium described above.
Alessandrini F., Ceresa D., Appolloni I., Marubbi D, & Malatesta P. (2016). Noninvasive Monitoring of Glioma Growth in the Mouse. Journal of Cancer, 7(13), 1791-1797.
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9

Serum-free Spheroid Formation Assay

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Cells were seeded at a density of 5×103 cells/well in 6-well ultra-low attachment plates (Corning Inc., Corning, NY) and cultured in serum-free DMEM:Ham's F12 medium (Invitrogen, Carlsbad, CA) containing 20 ng/ml human recombinant epidermal growth factor (PeproTech, Rocky Hill, NJ), 10 ng/ml human recombinant fibroblast growth factor-2 (PeproTech), 1% Insulin-Transferrin-Selenium Solution (ITS-G), 200 U/ml penicillin, and 100 µg/ml streptomycin. After 12 days, the number of spheres consisting of >20 cells was counted and imaged under a light microscope. The experiment was performed twice.
Fujiwara K., Ohuchida K., Sada M., Horioka K., Ulrich CD I.I.I., Shindo K., Ohtsuka T., Takahata S., Mizumoto K., Oda Y, & Tanaka M. (2014). CD166/ALCAM Expression Is Characteristic of Tumorigenicity and Invasive and Migratory Activities of Pancreatic Cancer Cells. PLoS ONE, 9(9), e107247.
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