Huvec cells

Manufactured by PromoCell

Sourced in Germany

HUVEC cells are primary human umbilical vein endothelial cells. They are isolated from the human umbilical vein and are commonly used in cell culture research to study endothelial cell biology.

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6 protocols using huvec cells

Cell Culture Protocols for Diverse Respiratory Cell Types

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Human A549-type II pneumocyte and murine RAW264.7 macrophage cell lines were obtained from the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany); human BEAS-2B bronchial epithelial cell line was obtained from Sigma-Aldrich (Taufkirchen, Germany) and primary human umbilical vein endothelial cells (HUVEC) from Pelobiotech (Planegg, Germany). Murine hematopoietic progenitor Hoxb8 neutrophils [19 (link)] were a generous gift from Prof. Georg Häcker (Institute of Medical Microbiology and Hygiene, UMC, Freiburg, Germany).
Cell cultures were grown under standard conditions using corresponding culture medium containing 1% penicillin-streptomycin (100 U/mL) as follows: A549 and RAW264.7 in DMEM-GlutaMAX supplemented with 10% fetal bovine serum (FBS; Gibco, Life Technologies, Paisley, Scotland), BEAS-2B in RPMI 1640-GlutaMAX (Gibco) supplemented with 5% FBS, primary HUVEC cells in Endothelial Cell Growth Medium Kit (PromoCell, Heidelberg, Germany), and Hoxb8 neutrophils in Opti-MEM-GlutaMAX (Gibco) supplemented with 10% heat-inactivated FBS, 30 µM 2-mercaptoethanol (Gibco), 1 µM estradiol and 1% stem cell factor. Prior to the experiments, progenitor Hoxb8 neutrophils were cultured for four days in differentiating medium (estradiol free medium), as described previously [19 (link)].

Spassov S.G., Faller S., Goeft A., von Itter M.N., Birkigt A., Meyerhoefer P., Ihle A., Seiler R., Schumann S, & Hoetzel A. (2022). Profiling Distinctive Inflammatory and Redox Responses to Hydrogen Sulfide in Stretched and Stimulated Lung Cells. Antioxidants, 11(5), 1001.

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Fluorescent Labeling of Cell Lines

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Fluorescein isothiocyanate (FITC-) and tetramethyl rhodamine isothiocyanate (TRITC-) labeled dextran dyes (70 kDa FITC-dextran), Alexa Fluor 555, and carbocyanine DiD were purchased from Invitrogen (Carlsbad, CA); TNF-α was purchased from Sigma (St. Louis, MO); anti- HSP20, HSP70, vWF, ICAM-1, and E-selectin (CD62E) were purchased from Abcam (Cambridge, MA). All organic solvents used were of analytical grade and used as received. Human cancer cell lines, SUM159 and MCF-7, were obtained from American Tissue Culture Collection (Manassas, VA) and HUVEC cells were obtained from PromoCell (La Jolla, CA). Cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) (Mediatech, Inc., Manassas, VA) supplemented with 10% (v/v) FBS (Sigma Aldrich, St. Louis, MO) while HUVECs were cultured in Endothelial cell culture medium (PromoCell, La Jolla, CA). All cell lines were grown at 37°C in a humidified incubator containing 5% CO₂.

Kirui D.K., Mai J., Palange A.L., Qin G., van de Ven A.L., Liu X., Shen H, & Ferrari M. (2014). Transient Mild Hyperthermia Induces E-selectin Mediated Localization of Mesoporous Silicon Vectors in Solid Tumors. PLoS ONE, 9(2), e86489.

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HUVEC Cell Culture and Rhamnazin Treatment

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HUVEC cells were purchased from PromoCell (Heidelberg, Germany) and cultured in EBM2 medium containing 2% fetal bovine serum and endothelial cell growth supplement (PromoCell) at 37 °C. Cell growth was observed by inverted microscope. The logarithmic phase endothelial cells were used during the experiment. Rhamnazin (98%, Sigma-Aldrich, St. Louis, MO) was dissolved in DMSO to prepare required concentration.

Yu Y., Zhou X.Z., Ye L., Yuan Q., Freeberg S., Shi C., Zhu P.W., Bao J., Jiang N, & Shao Y. (2018). Rhamnazin attenuates inflammation and inhibits alkali burn-induced corneal neovascularization in rats. RSC Advances, 8(47), 26696-26706.

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Dynamic Tumor Cell Adhesion Under Flow

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The dynamic adhesion of human HT29 and mouse MC38 tumor cells to endothelial cells under physiological flow conditions was analyzed in a laminar flow adhesion assay as previously described.⁷⁰ (link) In short, HUVEC cells (PromoCell, Heidelberg) or LSECs were seeded in ibidiTreat μ-slide IV0.4 (ibidi GmbH, Germany) flow chambers and after confluence to monolayers, endothelial cells were stimulated with 10 ng/mL recombinant human or murine IL-1α (R&D Systems, Minneapolis, MN), 10 ng/mL recombinant human or murine TNFα or 20 to 100 ng/mL human or murine IL-22 for 4 h prior to the flow assay (tumor cell suspension: 1x10⁵ cells/ml; flow rate: 8.5 mL/h; shear stress: 0.25 dyn/cm²). Non-stimulated endothelial cells served as negative controls. Data were acquired and evaluated with CapImage software (version 8.6, Dr. Heinrich Zeintl, Heidelberg). The adhesive events on the endothelium were distinguished into firm adhesion, rolling and tethering and counted in three separate regions of interest for 1 min each.

Giannou A.D., Kempski J., Shiri A.M., Lücke J., Zhang T., Zhao L., Zazara D.E., Cortesi F., Riecken K., Amezcua Vesely M.C., Low J.S., Xu H., Kaffe E., Garcia-Perez L., Agalioti T., Yamada Y., Jungraithmayr W., Zigmond E., Karstens K.F., Steglich B., Wagner J., Konczalla L., Carambia A., Schulze K., von Felden J., May P., Briukhovetska D., Bedke T., Brockmann L., Starzonek S., Lange T., Koch C., Riethdorf S., Pelczar P., Böttcher M., Sabihi M., Huber F.J., Reeh M., Grass J.K., Wahib R., Seese H., Stüben B.O., Fard-Aghaie M., Duprée A., Scognamiglio P., Plitzko G., Meiners J., Soukou S., Wittek A., Manthey C., Maroulis I.C., Arck P.C., Perez D., Gao B., Zarogiannis S.G., Strowig T., Pasqualini R., Arap W., Gosálvez J.S., Kobold S., Prinz I., Guse A.H., Tachezy M., Ghadban T., Heumann A., Li J., Melling N., Mann O., Izbicki J.R., Pantel K., Schumacher U., Lohse A.W., Flavell R.A., Gagliani N, & Huber S. (2023). Tissue resident iNKT17 cells facilitate cancer cell extravasation in liver metastasis via interleukin-22. Immunity, 56(1), 125-142.e12.

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Biofunctionalized Polymer Coatings for Cell Culture

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CuBr₂ (99.999% trace metal basis), CuBr (99.99%), 2,2’-bipyridyl (99%), oligo(ethylene glycol) methyl ether methacrylate (M_n = 300 g/mol, MeOEGMA), glycidyl methacrylate (97%, GMA), ethylenediamine (99.5%, EtDA), α-bromoisobutyryl bromide (98%, BiBB), copper sulphate (CuSO₄), sodium L-ascorbate (98%), NaN₃ (99.5%) were purchased from Sigma-Aldrich (Czech Republic). Deionized (DI) water was obtained from a Milli-Q purification system (Milli-Q gradient A10, Merck-Millipore). Tetrahydrofuran (THF, Lach-Ner, Czech Republic) was distilled under argon over sodium immediately prior use. All other organic solvents (Lach-Ner, Czech Republic) of analytical grade were used as received. ATRP initiators 11-(2-bromo-2-methyl)propionyloxy)undecyltrichlorosilane and ω-mercaptoundecyl bromoisobutyrate were synthesized according to procedures reported earlier [49 (link),50 (link)]. Human blood plasma, Endothelial Cell Growth Medium 2 Kit, and HUVEC cells were obtained from Promocell (Germany). Penicillin-streptomycin (10 U/mL, 10 μg/mL), trypsin-EDTA (0.05%–0.02%) and Phalloidin–Atto 594 were from Sigma-Aldrich (Czech Republic). LIVE/DEAD™ Viability/Cytotoxicity Kit for mammalian cells and Hoechst 33342 were purchased from Thermo Fisher Scientific (USA).

Sivkova R., Táborská J., Reparaz A., de los Santos Pereira A., Kotelnikov I., Proks V., Kučka J., Svoboda J., Riedel T, & Pop-Georgievski O. (2020). Surface Design of Antifouling Vascular Constructs Bearing Biofunctional Peptides for Tissue Regeneration Applications. International Journal of Molecular Sciences, 21(18), 6800.

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Establishing Pancreatic and Bone Cancer Cell Lines

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MiaPaCa-2 (CRL-1420), BxPC3 (CRL-1687), Capan-1 (HTB-79), Panc-1 (CRL-1469) and U2OS (HTB-96) cell lines were purchased from the American Type Culture Collection (Rockville, MD, USA) and were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal calf serum (FCS), 2 mM L-glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin (Gibco, Saint-Aubin, France) in a 5% CO₂ humidified incubator at 37 °C. HUVEC cells were obtained from PromoCell GmbH (Heidelberg, Germany). U2OS cell line stably expressing human apelin receptor (hAPJ) and β-arrestin2-GFP were already described [31 (link)]. No mycoplasma-positive cells were used in this work.
To transiently express the dominant negative K44A mutant of Dynamin, MiaPaCa-2 cells were transfected using Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, MA, USA). MiaPaCa-2 cells expressing control hairpins or hairpins targeting APJ were transduced with lentiviral, encoding for secreted lucia luciferase (Invivogen), as described previously [57 (link)].

Chaves-Almagro C., Auriau J., Dortignac A., Clerc P., Lulka H., Deleruyelle S., Projetti F., Nakhlé J., Frances A., Berta J., Gigoux V., Fourmy D., Dufresne M., Gomez-Brouchet A., Guillermet-Guibert J., Cordelier P., Knibiehler B., Jockers R., Valet P., Audigier Y, & Masri B. (2022). Upregulated Apelin Signaling in Pancreatic Cancer Activates Oncogenic Signaling Pathways to Promote Tumor Development. International Journal of Molecular Sciences, 23(18), 10600.

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