Human recombinant egf

Manufactured by STEMCELL

Sourced in Canada

Human recombinant EGF is a protein that promotes cell growth and proliferation. It is a commonly used cell culture supplement for in vitro studies.

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Lab products found in correlation

Heparin, by STEMCELL (5 mentions) Human recombinant bfgf, by STEMCELL (5 mentions) Basic fibroblast growth factor (bfgf), by STEMCELL (2 mentions) Neurocult ns a proliferation kit, by STEMCELL (2 mentions) Heparin solution, by STEMCELL (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Recombinant human bfgf, by Miltenyi Biotec (1 mentions) Rlt lysis buffer, by Qiagen (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Complete differentiation medium, by STEMCELL (1 mentions) Neurocult proliferation kit mouse and rat, by STEMCELL (1 mentions) Matrigel, by Roche (1 mentions) Laminin, by Merck Group (1 mentions) Neurocult nsc basal medium mouse, by STEMCELL (1 mentions) Neurocult mouse proliferation supplement, by STEMCELL (1 mentions) Matrigel, by Corning (1 mentions) Accutase, by STEMCELL (1 mentions) Phospho akt s473, by Cell Signaling Technology (1 mentions) Phospho stat3 y705, by Cell Signaling Technology (1 mentions)

12 protocols using human recombinant egf

Sphere Culture Protocols for Cell Expansion

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For sphere cultures, cells were seeded at a low density (3000 viable cells/cm²) onto non-adherent plates and cultured in serum-free DMEM/F-12 medium, supplemented with 2% B27 (Gibco Life Technologies, Rockville, MD, USA), 20 ng/mL recombinant human bFGF (Miltenyi Biotec, Bergisch Gladbach, Germany), 20 ng/mL recombinant human EGF (StemCell Technologies, Vancouver, BC, Canada), 0.0002% heparin (StemCell Technologies, Vancouver, BC, Canada) and antibiotics (100 U/mL penicillin, 100 µg/mL streptomycin, Gibco Life Technologies, Rockville, MD, USA). Every 3 days, 25% of the medium was replaced. After 7–14 days of culturing, the spheres were collected by centrifugation at 110× g and lysed in Qiagen RLT lysis buffer for RNA isolation or lysed in buffer supplemented with complete protease inhibitor cocktail (Roche Applied Science, Indianapolis, IN, USA) for blotting.

Ciechomska I.A., Wojnicki K., Wojtas B., Szadkowska P., Poleszak K., Kaza B., Jaskula K., Dawidczyk W., Czepko R., Banach M., Czapski B., Nauman P., Kotulska K., Grajkowska W., Roszkowski M., Czernicki T., Marchel A, & Kaminska B. (2023). Exploring Novel Therapeutic Opportunities for Glioblastoma Using Patient-Derived Cell Cultures. Cancers, 15(5), 1562.

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Derivation of Neural Lineage Cells

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Neural stem cells (NSCs) were harvested from the subventricular zone of 8-12 week old mice and grown as neurospheres in suspension culture as described previously [42 ]. Proliferating cultures were maintained in Neurocult complete proliferation medium, which is comprised of NeuroCult proliferation kit mouse and rat (StemCell Technologies) supplemented with 10 ng/mL recombinant human FGF (StemCell Technologies), 20 ng/mL recombinant human EGF (StemCell Technologies) and 2 μg/mL heparin (StemCell Technologies). To produce monolayer cultures of neural lineage cells (neurons, astrocytes and oligodendrocytes), NSCs were plated as single cell suspensions onto Matrigel (Roche) coated vessels and incubated in complete differentiation medium (StemCell Technologies) for 7 days, as described previously [43 (link)].

Foliaki S.T., Wood A., Williams K., Smith A., Walters R.O., Baune C., Groveman B.R, & Haigh C.L. (2023). Temporary alteration of neuronal network communication is a protective response to redox imbalance that requires GPI-anchored prion protein. Redox Biology, 63, 102733.

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Derivation and Culture of Mouse Neural Stem Cells

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NSCs were derived and cultured exactly as previously described^{13 (link),14 (link)}. In brief, cortices of E14.5 embryos were dissociated and grown as an adherent monolayer on tissue culture-treated plates. Growth media was NeuroCult NSC Basal Medium (Mouse) containing NeuroCult Proliferation Supplement (Mouse) (StemCell Technologies), recombinant human EGF (20 ng/mL), recombinant human basic FGF (20 ng/mL, all from Stem Cell Technologies) and 10 ng/ml laminin (Sigma Aldrich). Primary cells were maintained for a maximum of 20 passages.

Tapia del Fierro A., den Hamer B., Benetti N., Jansz N., Chen K., Beck T., Vanyai H., Gurzau A.D., Daxinger L., Xue S., Ly T.T., Wanigasuriya I., Iminitoff M., Breslin K., Oey H., Krom Y.D., van der Hoorn D., Bouwman L.F., Johanson T.M., Ritchie M.E., Gouil Q.A., Reversade B., Prin F., Mohun T., van der Maarel S.M., McGlinn E., Murphy J.M., Keniry A., de Greef J.C, & Blewitt M.E. (2023). SMCHD1 has separable roles in chromatin architecture and gene silencing that could be targeted in disease. Nature Communications, 14, 5466.

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Organoid Colony Formation Assay

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Colony formation assays were performed according to the manufacturer’s protocol (Stem Cell Technologies, 05610). Freshly sorted tdTomato⁺ and tdTomato^- cells were embedded in 50% Matrigel (Corning, 356231) and cultured in mouse Epicult-B medium (Stem Cell Technology, 05610) containing 5% FBS, 20 ng/mL Human Recombinant EGF (Stem Cell Technology, 78006.1), 20 ng/mL Human Recombinant bFGF (Stem Cell Technology, 78003.1), and 4 μg/mL heparin solution (Stem Cell Technology, 07980). FACS-sorted cells were resuspended at a density of 80,000 cells/mL in chilled 100% growth-factor-reduced Matrigel (BD Bioscience). The medium was changed every 2 days, and colonies were scored after 7∼8 days. Colony size was analyzed with Adobe Photoshop CC 2018.

Liu R., Hu H., McNeil M., Xu J., Bi X., Lou P., Guerrero-Juarez C.F., Dai X., Plikus M.V., Shuai J., Yu Z, & Lv C. (2022). Dormant Nfatc1 reporter-marked basal stem/progenitor cells contribute to mammary lobuloalveoli formation. iScience, 25(3), 103982.

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Characterization of Glioblastoma Cell Lines

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293T (source: female), human glioma cell line U87 cells (source: male) were cultured in DMEM (Hyclone) with 10%
heat-inactivated FBS (Hyclone), 100 U/ml Penicillin and 100 μg/ml Streptomycin (Gibico/Life Technologies). GBM stem
cell lines TS528, TS543, TS576 and TS600 were kindly provided by Dr. Cameron Brennan, MSKCC; HK281 was from Dr. Harley
Kornblum, UCLA; GSC11 (source: male) and GSC23 (source: male) were acquired from Dr. Frederick Lang, MD Anderson; GBM6
(source: male) and GBM39 (source: male) were obtained from Dr. David James, Northwestern University; GBM1600 (source: male)
was kindly provided by Dr. Paul Mischel, Ludwig Institute. GBM stem cell (Neurosphere) lines were cultured in DMEM/F12 medium
supplemented with B27 (GIBCO/Life Technologies) and human recombinant EGF (20 ng/ml), bFGF (20 ng/ml) and 2 mg/mL heparin
(StemCell Technologies). Neurospheres were dissociated with Accutase (Stemcell Technologies). Primary neural stem cells were
isolated from the brain of E16.5 littermates of WT and Y240F PTEN mice. Cells were cultured in the same medium as GBM stem
cell lines. All cells were kept in a humidified 5% CO2 incubator at 37°C.

Ma J., Benitez J.A., Li J., Miki S., Ponte de Albuquerque C., Galatro T., Orellana L., Zanca C., Reed R., Boyer A., Koga T., Varki N.M., Fenton T.R., Marie S.K., Lindahl E., Gahman T.C., Shiau A.K., Zhou H., DeGroot J., Sulman E.P., Cavenee W.K., Kolodner R.D., Chen C.C, & Furnari F.B. (2019). Inhibition of nuclear PTEN tyrosine phosphorylation enhances glioma radiation sensitivity through attenuated DNA repair. Cancer cell, 35(3), 504-518.e7.

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Glioblastoma Cell Culturing and Characterization

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U87 glioma cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% FBS and 1% penicillin/streptomycin. HK281 GBM-spheres [a kind gift from Harley Kornblum, UCLA, (Visnyei et al., 2011 (link))] were cultured in DMEM/F12 medium supplemented with 1× B27 (GIBCO/Life Technologies), 1% penicillin/streptomycin, human recombinant EGF (20 ng/mL), bFGF (20 ng/mL) and 2 mg/mL heparin (StemCell Technologies). All cells were maintained at 37°C, 5% CO₂, and 100% relative humidity for the duration of the experiment. Reagents used in this study were obtained from the following sources: Antibodies: EGFR (BD Biosciences), pan-phospho-Tyrosine-HRP (R&D Systems), p42/44 MAPK, phospho-p42/44 MAPK, STAT3, phospho-STAT3 y705, AKT, phospho-AKT s473 (Cell Signaling Technology), Ki67 (Santa Cruz Biotechnology), and β-Actin (Sigma). Inhibitors: U0126 and PD98059 (LC Laboratories). The monoclonal antibody 806 was produced in the Biological Production Facility at the Olivia-Newton John Cancer Research Institute (Melbourne, Australia).

Binder Z.A., Thorne A.H., Bakas S., Wileyto E.P., Bilello M., Akbari H., Rathore S., Ha S.M., Zhang L., Ferguson C.J., Dahiya S., Bi W.L., Reardon D.A., Idbaih A., Felsberg J., Hentschel B., Weller M., Bagley S.J., Morrissette J.J., Nasrallah M.P., Ma J., Zanca C., Scott A.M., Orellana L., Davatzikos C., Furnari F.B, & O’Rourke D.M. (2018). Epidermal Growth Factor Receptor Extracellular Domain Mutations in Glioblastoma Present Opportunities for Clinical Imaging and Therapeutic Development. Cancer cell, 34(1), 163-177.e7.

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Isolation and Culture of GCSCs and PCSCs

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Fresh surgical GBM and peritumoral specimens (at a distance <1 cm from macroscopic border), derived from five patients, were dissected and digested in papain solution (Worthington Biochemical, Lakewood, NJ, USA). The neurosphere cells, termed in this work Glioblastoma Cancer Stem Cells (GCSCs) and Peritumoral Cancer Stem Cells (PCSCs) were isolated, cultured and maintained in NeuroCult™ NS-A Proliferation Kit (Stemcell Technologies Inc, Vancouver, BC, Canada) supplemented with 20 ng/ml human recombinant EGF, 10 ng/ml human recombinant bFGF and 2 ug/ml heparin (all from StemCell Technologies Inc.), as described previously [47 (link), 48 (link)]. Commercially available pooled Human Umbilical Vein Endothelial Cells (HUVEC) were obtained from Lonza Sales Ldt (Switzerland), cultured in EGM-2 Endothelial Cell Growth Medium-2 (Endothelial Basal Medium EBM-2 + EGM-2 Bullet Kit, Lonza), supplemented with 100 units/ml penicillin/streptomycin (Life Technologies Corporation, Gaithersburg, MD, USA). U87MG grade IV glioma cell line was kindly provided by Dr. Emilio Ciusani, (“Carlo Besta” National Neurological Institute, Milan, Italy) and maintained in DMEM containing 10% (v/v) fetal calf serum, 200 mM L-glutamine, 100 units/ml penicillin/streptomycin (Life Technologies). All cell types were maintained at 37°C in a 5% CO₂ humidified atmosphere.

D'Alessio A., Proietti G., Lama G., Biamonte F., Lauriola L., Moscato U., Vescovi A., Mangiola A., Angelucci C, & Sica G. (2016). Analysis of angiogenesis related factors in glioblastoma, peritumoral tissue and their derived cancer stem cells. Oncotarget, 7(48), 78541-78556.

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Hindbrain Cell Culture Protocol

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Cell cultures from E10.5 hindbrains were prepared and imaged as described above, with the following modifications: hindbrains were collected in PBS containing calcium and magnesium (Biological Industries, Israel), and cells were grown in hESC media combined with NeuroCult Proliferation Supplement, supplemented with 20 μg of human recombinant EGF, 10 μg human recombinant bFGF and 10 μg 0.2% heparin solution (all from STEMCELL Technologies).

Hutchings C., Nuriel Y., Lazar D., Kohl A., Muir E., Genin O., Cinnamon Y., Benyamini H., Nevo Y, & Sela-Donenfeld D. (2024). Hindbrain boundaries as niches of neural progenitor and stem cells regulated by the extracellular matrix proteoglycan chondroitin sulphate. Development (Cambridge, England), 151(4), dev201934.

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Glioblastoma Cancer Stem Cells Characterization

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A total of six pairs of neurospheres derived from both GBM and peritumoral tissue (at a distance < 1 cm from macroscopic tumor border), called Glioblastoma Cancer Stem Cells (GCSCs) and Peritumoral Cancer Stem Cells (PCSCs), respectively, were kindly provided by Vescovi and Binda. According to the classification of human gliomas by WHO [29 (link)], we analyzed the GCSC and PCSC neurospheres for the mutational status of IDH1 [30 (link), 31 (link)]. Mutational IDH1/2 status and MGMT promoter methylation status were also investigated in all the available tissue samples from which the neurospheres were derived [32 (link)].
The neurospheres were cultured in NeuroCult™ NS-A Proliferation Kit (StemCell Technologies Inc, Vancouver, BC, Canada) supplemented with 20 ng/mL human recombinant EGF, 10 ng/ml human recombinant bFGF and 2 µg/mL heparin (all from StemCell Technologies Inc.), as described previously [33 (link)]. All cell cultures were maintained at 37 °C in a 5% CO2 humidified atmosphere.

Scicchitano B.M., Sorrentino S., Proietti G., Lama G., Dobrowolny G., Catizone A., Binda E., Larocca L.M, & Sica G. (2018). Levetiracetam enhances the temozolomide effect on glioblastoma stem cell proliferation and apoptosis. Cancer Cell International, 18, 136.

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Propagation and Culturing of Patient-Derived Glioma Cell Lines

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Human patient-derived glioma cell lines GBM6, GBM12, and GBM39 were propagated in mouse flanks before use in culture [46 (link)]. GBM6 and GBM39 spheroids were generated in human neurobasal media (human Neurocult NS-A Basal Medium; Cat 5750; StemCell Technologies, Vancouver, Canada) supplemented with 20 ng/mL human recombinant EGF (Cat 78006; StemCell Technologies), 10 ng/mL human recombinant beta-FGF (Cat 78003; StemCell Technologies), 0.5% StemPro Neural Supplement proliferation media [Cat A1050801; Thermofisher Scientific), and 1% penicillin/streptomycin). Cells were grown in a humidified incubator at 37 °C and 5% CO₂.

Pawlowski K.D., Duffy J.T., Tiwari A., Zannikou M, & Balyasnikova I.V. (2023). Bi-Specific Killer Cell Engager Enhances NK Cell Activity against Interleukin-13 Receptor Alpha-2 Positive Gliomas. Cells, 12(13), 1716.

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