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Human recombinant basic fibroblast growth factor

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Human recombinant basic fibroblast growth factor is a laboratory reagent used to stimulate cell growth and proliferation. It is a protein that acts as a growth factor, promoting the development and survival of various cell types. This product is intended for research use only and its detailed applications should be determined by the user.

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Lab products found in correlation

Glutamax, by Thermo Fisher Scientific (5 mentions) Chemically defined lipid concentrate, by Thermo Fisher Scientific (4 mentions) Recombinant human epidermal growth factor, by R&D Systems (4 mentions) Insulin transferrin selenium, by Thermo Fisher Scientific (4 mentions) Penicillin streptomycin antibiotic, by Merck Group (4 mentions) Mcdb 131 medium, by Thermo Fisher Scientific (4 mentions) Hepes, by Merck Group (4 mentions) Ascorbic acid, by Merck Group (4 mentions) Hydrocortisone, by Merck Group (4 mentions) Human recombinant epidermal growth factor, by Merck Group (2 mentions) Insulin, by Merck Group (2 mentions) Dmem f12 medium, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Ultra low attachment 24 well culture plates, by Corning (1 mentions) Trypsin inhibitor, by Thermo Fisher Scientific (1 mentions) Tryple express, by Thermo Fisher Scientific (1 mentions) Methylcellulose, by Merck Group (1 mentions) Epidermal growth factor (egf), by Merck Group (1 mentions) Epithelial growth factor (egf), by Merck Group (1 mentions)

Spelling variants of this product

Human basic fibroblast growth factor (44 citations) Fibroblast growth factor-basic (2 citations)

10 protocols using human recombinant basic fibroblast growth factor

1

Isolation and Culture of HUVECs

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Cells were harvested from fresh umbilical cords obtained during normal delivery of healthy neonates (according to Helsinki Protocol, Semmelweis University Institutional Review Board specifically approved this study, (permission number: SETUKEB 141/2015), and all participants provided their written informed consent to participate in this study), by collagenase digestion as described earlier.42 ,43 HUVECs were kept in gelatin-precoated flasks in MCDB131 medium (Life Technologies, Carlsbad, CA, USA) completed with 5% heat-inactivated FCS, 2 ng/ml human recombinant epidermal growth factor (R&D Systems), 1 ng/ml human recombinant basic fibroblast growth factor (Sigma), 0.3% Insulin transferrin selenium (Life Technologies), 1% chemically defined lipid concentrate (Life Technologies), 1% glutamax (Life Technologies), 1% penicillin–streptomycin antibiotics (Sigma), 5 μg/ml ascorbic acid (Sigma), 250 nM hydrocortisone (Sigma), 10 mM HEPES (Sigma), and 7.5 U/ml heparin hereinafter referred to as Comp-MCDB131. Each experiment was performed on at least 3 independent primary HUVEC cultures from different individuals.
Kolonics F., Kajdácsi E., Farkas V.J., Veres D.S., Khamari D., Kittel Á., Merchant M.L., McLeish K.R., Lőrincz Á.M, & Ligeti E. (2020). Neutrophils produce proinflammatory or anti-inflammatory extracellular vesicles depending on the environmental conditions. Journal of leukocyte biology, 109(4), 793-806.
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2

Enrichment and Characterization of Cancer Stem Cells

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For enrichment of CSCs, cancer cells were seeded into ultra-low attachment 24-well culture plates (Corning) at a density of 20,000 cells/well and cultured in serum-free DMEM/ F12 medium (Gibco), containing 20 ng/mL human recombinant epidermal growth factor (Sigma-Aldrich), 20 ng/mL human recombinant basic fibroblast growth factor (Sigma-Aldrich), 1:50 dilution of B27 (Gibco) and 5 μg/mL insulin (Sigma-Aldrich). After 5 ~ 7 days of culture, the cells formed CSCs-like aggregates and the number of tumor spheres was counted under microscope.
Guo M., Lian J., Liu Y., Dong B., He Q., Zhao Q., Zhang H., Qi Y., Zhang Y, & Huang L. (2022). Loss of miR-637 promotes cancer cell stemness via WASH/IL-8 pathway and serves as a novel prognostic marker in esophageal squamous cell carcinoma. Biomarker Research, 10, 77.
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3

Culturing and Propagating Spheres from Single Cells

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Single cells were cultured in 300 μl of serum-free DMEM/F12 medium (Invitrogen) supplemented with 20 ng/ml human recombinant epidermal growth factor (Sigma-Aldrich), 10 ng/ml human recombinant basic fibroblast growth factor (Sigma-Aldrich), 4 μg/ml insulin (Sigma-Aldrich), B27 (1:50; Invitrogen), 500 U/ml penicillin, 500 μg/ml streptomycin (Invitrogen) and 1% methylcellulose (Sigma-Aldrich). Cells were cultured in suspension in poly-HEMA-coated 24-well plates. Cells were replenished with 30 μl of supplemented medium every second day. To propagate spheres in vitro, spheres were collected by gentle centrifugation and dissociated to single cells using TrypLE Express (Invitrogen). Following dissociation, trypsin inhibitor (Invitrogen) was used to neutralize the reaction, and the cells were cultured to generate the next generation of spheres.
Ng K.Y., Chai S., Tong M., Guan X.Y., Lin C.H., Ching Y.P., Xie D., Cheng A.S, & Ma S. (2016). C-terminal truncated hepatitis B virus X protein promotes hepatocellular carcinogenesis through induction of cancer and stem cell-like properties. Oncotarget, 7(17), 24005-24017.
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4

Isolation and Culture of HUVECs

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Cells were harvested from fresh umbilical cords obtained during normal delivery of healthy neonates (according to Helsinki Protocol, Semmelweis University Institutional Review Board specifically approved this study, (permission number: SETUKEB 141/2015), and all participants provided their written informed consent to participate in this study), by collagenase digestion as described earlier.42 ,43 HUVECs were kept in gelatin-precoated flasks in MCDB131 medium (Life Technologies, Carlsbad, CA, USA) completed with 5% heat-inactivated FCS, 2 ng/ml human recombinant epidermal growth factor (R&D Systems), 1 ng/ml human recombinant basic fibroblast growth factor (Sigma), 0.3% Insulin transferrin selenium (Life Technologies), 1% chemically defined lipid concentrate (Life Technologies), 1% glutamax (Life Technologies), 1% penicillin–streptomycin antibiotics (Sigma), 5 μg/ml ascorbic acid (Sigma), 250 nM hydrocortisone (Sigma), 10 mM HEPES (Sigma), and 7.5 U/ml heparin hereinafter referred to as Comp-MCDB131. Each experiment was performed on at least 3 independent primary HUVEC cultures from different individuals.
Kolonics F., Kajdácsi E., Farkas V.J., Veres D.S., Khamari D., Kittel Á., Merchant M.L., McLeish K.R., Lőrincz Á.M, & Ligeti E. (2020). Neutrophils produce proinflammatory or anti-inflammatory extracellular vesicles depending on the environmental conditions. Journal of leukocyte biology, 109(4), 793-806.
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5

Isolation and Culture of HUVECs

Cited 1 time
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Cells were prepared from fresh umbilical cords obtained during normal deliveries of healthy neonates1 (link), 29 (link). HUVECs were grown in gelatin-precoated flasks (Corning® Costar®) in MCDB131 medium (Life Technologies) completed with 5% heat-inactivated fetal calf serum (FCS), 2 ng/ml human recombinant epidermal growth factor (R&D Systems), 1 ng/ml human recombinant basic fibroblast growth factor (Sigma), 0.3% Insulin Transferrin Selenium (Life Technologies), 1% Chemically Defined Lipid Concentrate (Life Technologies), 1% Glutamax (Life Technologies), 1% Penicillin-Streptomycin antibiotics (Sigma), 5 µg/ml Ascorbic acid (Sigma), 250 nM Hydrocortisone (Sigma), 10 mM Hepes (Sigma), and 7.5 U/ml Heparin. Each experiment was performed on primary HUVEC cultures from different individuals before the 4th passage. The study was conducted in conformity with the WMA Declaration of Helsinki; its protocol was approved by the Semmelweis University Institutional Review Board (permission number: TUKEB64/2008), and all participants provided their written informed consent before inclusion.
Schwaner E., Németh Z., Jani P.K., Kajdácsi E., Debreczeni M.L., Doleschall Z., Dobó J., Gál P., Rigó J., András K., Hegedűs T, & Cervenak L. (2017). Transcriptome analysis of inflammation-related gene expression in endothelial cells activated by complement MASP-1. Scientific Reports, 7, 10462.
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6

Sphere Formation Assay for Cancer Stem Cells

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The sorted SP cells and non-SP cells were plated at a density of 1,000 cells/ml and suspended in tumor sphere medium consisting of serum-free 1:1 mixture of Ham's F-12/DMEM, N2 supplement, 10 ng/ml human recombinant basic fibroblast growth factor (Sigma-Aldrich) and 10 ng/ml epidermal growth factor(Sigma-Aldrich), and subsequently cultured in ultra-low attachment plates for ~2 weeks. Sorted SP and non-SP cells were seeded at a low density of 20 cells/l and the number of generated spheres (>100 μM) was counted following 7 days of culture. The values presented in the graph are the mean values of three independent experiments.
CHEN W., ZHANG Y.W., LI Y., ZHANG J.W., ZHANG T., FU B.S., ZHANG Q, & JIANG N. (2016). Constitutive expression of Wnt/β-catenin target genes promotes proliferation and invasion of liver cancer stem cells. Molecular Medicine Reports, 13(4), 3466-3474.
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7

Isolation and Culture of HUVECs

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Cells were harvested from fresh umbilical cords obtained during normal deliveries of healthy neonates by collagenase digestion as described earlier [13, 30] . HUVECs were kept in gelatin-precoated flasks (Corning® Costar®) in MCDB131 medium (Life Technologies) completed with 5% heat-inactivated fetal calf serum (FCS), 2 ng/ml human recombinant epidermal growth factor (R&D Systems), 1 ng/ml human recombinant basic fibroblast growth factor (Sigma), 0.3% Insulin Transferrin Selenium (Life Technologies), 1% Chemically Defined Lipid Concentrate (Life Technologies), 1% Glutamax (Life Technologies), 1% Penicillin-Streptomycin antibiotics (Sigma), 5 µg/ml Ascorbic acid (Sigma), 250 nM Hydrocortisone (Sigma), 10 mM Hepes (Sigma), and 7.5 U/ml Heparin (this completed medium is hereinafter: Comp-MCDB). Each experiment was performed on at least three independent primary HUVEC cultures from different individuals before the 4th passage. The study was conducted in conformity with the WMA Declaration of Helsinki; its protocol was approved by the Semmelweis University Institutional Review Board (permission number: TUKEB64/2008), and all participants provided their written informed consent before inclusion.
Jani P.K., Schwaner E., Kajdácsi E., Debreczeni M.L., Ungai-Salánki R., Dobó J., Doleschall Z., Rigó J J.r., Geiszt M., Szabó B., Gál P, & Cervenak L. (2016). Complement MASP-1 enhances adhesion between endothelial cells and neutrophils by up-regulating E-selectin expression. Molecular immunology, 75.
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8

Culturing Diverse Cell Lines

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The bovine macrophage cell line (BoMac), the human microglia cell line (HMC‐3), and the human epithelial colorectal adenocarcinoma cell line (Caco‐2) were grown in Dulbecco's modified Eagle's medium (DMEM) with Glutamax (Life Technologies, Zug, Switzerland) supplemented with 10% fetal calf serum (FCS) (Bioswisstec, Schaffhausen, Switzerland), 100U/ml penicillin and 10 µg/ml streptomycin (Life Technologies). Fetal bovine brain cells (FBBC‐1) were grown in a DMEM/F12 mix (1:1, Life Technologies) supplemented with 10% FCS, 50 ng/ml epithelial growth factor, 50 ng/ml recombinant human basic fibroblast growth factor (bFGF) (Sigma‐Aldrich, Buchs, Switzerland), 100 U/ml penicillin, 10 µg/ml streptomycin, and 1x N2 supplement (Life Technologies).
Gözel B., Monney C., Aguilar‐Bultet L., Rupp S., Frey J, & Oevermann A. (2019). Hyperinvasiveness of Listeria monocytogenes sequence type 1 is independent of lineage I‐specific genes encoding internalin‐like proteins. MicrobiologyOpen, 8(7), e00790.
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9

Molecular Cloning and Expression Analysis

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All chemicals and media were purchased from Sigma, Chemical Co. (St Louis, MO, USA). The primers used in this study were synthesized by Invitrogen Technology (Beijing, China). PCR reactions were performed using Taq DNA Polymerase (Takara, Cat#: DR001A, Dalian, China). Anti-Growth Hormone Polyclonal Antibody was purchased from Thermo Scientific Biotechnology (Cat#: PA1-85521, Waltham, MA USA). Polyclonal goat Anti-mouse IgG/HRPwas purchased from Santa Cruz Biotechnology (Cat#: 330, CA USA). Rabbit polyclonal antibody for beta Actin Loading Control (Cat#: PM053-7) was purchased from MBL. Dulbecco’s Modified Eagle’s medium (DMEM, Cat#: 12800-017) AND G418 (Cat#: 11811031) was purchased from Invitrogen (Grand Island, NY USA). The serum was purchased from Hyclone (Cat#: SV30087.02, Logan, UT, USA). Recombinant human basic fibroblast growth factor (bFGF) was purchased from Sigma (Cat#: F0291). Endo Free plasmid mini-prep kits were purchased from Omega Bio-Tek (Cat#: D6950, Norcross, GA USA). The restriction enzymes were purchased from Fermentas (Ontario, Canada).
Ju H., Zhang J., Bai L., Mu Y., Du Y., Yang W., Li Y., Sheng A, & Li K. (2015). The transgenic cloned pig population with integrated and controllable GH expression that has higher feed efficiency and meat production. Scientific Reports, 5, 10152.
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10

Endothelial Cell Barrier Regulation

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Recombinant human basic fibroblast growth factor (bFGF) was purchased from Sigma (Sigma-Aldrich, St. Louis, MO), and PI3K inhibitor LY294002 (Sigma-Aldrich, St Louis, MO) was dissolved in 25 % dimethylsulfoxide solution (DMSO). FITC-dextran and Evans Blue were purchased from Sigma-Aldrich, St. Louis, MO. Anti-β-catenin, anti-p120-catenin, and anti-CD31 were purchased from Abcam, Cambridge, MA. GTP-Rac1, total-Rac1, GTP-RhoA, and total-RhoA were detected using Rac1/Cdc42 and Rho Activation Assay Kits (Millipore, Temecula, CA). Anti-Akt and anti-p-Akt (Ser473), anti-claudin-5, anti-occludin, and anti-zonula occludens-1 were from Proteintech. Human brain microvascular endothelial cells (HBMECs) and endothelial cell medium (ECM) were purchased from Sciencell (Carlsbad, CA, USA). The appropriate secondary antibodies were obtained from Santa Cruz Biotechnology.
Wang Z.G., Cheng Y., Yu X.C., Ye L.B., Xia Q.H., Johnson N.R., Wei X., Chen D.Q., Cao G., Fu X.B., Li X.K., Zhang H.Y, & Xiao J. (2015). bFGF Protects Against Blood-Brain Barrier Damage Through Junction Protein Regulation via PI3K-Akt-Rac1 Pathway Following Traumatic Brain Injury. Molecular neurobiology, 53(10), 7298-7311.
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