Huvecs

Manufactured by Addgene

HUVECs (Human Umbilical Vein Endothelial Cells) are primary cells derived from the umbilical vein of human newborns. They are used as an in vitro model to study endothelial cell biology and function.

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Lab products found in correlation

Huvecs, by Lonza (2 mentions) Egm 2 media, by Lonza (1 mentions) Mda mb 231 breast, by American Type Culture Collection (1 mentions) S2510, by Thermo Fisher Scientific (1 mentions) Superscript 4 vilo master mix, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Phosstop phosphatase inhibitor cocktail, by Roche (1 mentions)

2 protocols using huvecs

Culturing and Labeling Cell Lines

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Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza and cultured in EGM-2 media (Lonza, Switzerland). MDA-MB-231 breast cancer cells were purchased from ATCC and cultured in DMEM (low glucose) + 10% Fetal bovine serum (FBS) + 2 mM L-glutamine + 50 µg/ml Gentamycin. Human mesenchymal stem cells (MSCs, bone marrow-derived) and human lung fibroblasts (HLFs) were purchased from Lonza and cultured in DMEM (low glucose) + 10% FBS + 2 mM L-glutamine + 50 µg/ml Gentamycin. HUVECs were used at passages 3–8, and the stromal cells (MSCs and HLFs) were used at passages 3–6. Green Fluorescent Protein-labelled MDA-MB-231 cells (GFP-MDA-MB-231) were prepared by transducing MDA-MB-231 cells with a lentiviral construct pCSCG-EGFP (Addgene). Similarly, Red Fluorescent-Protein labelled HUVECs (mApple HUVECs) were prepared by transducing HUVECs with a lentiviral construct pBAD-mApple (Addgene). All the cells were maintained in standard tissue culture incubators at 37 °C, 95% humidity, and 5% CO₂.

Kwak T.J, & Lee E. (2020). In vitro modeling of solid tumor interactions with perfused blood vessels. Scientific Reports, 10, 20142.

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HUVEC Cell Culture and Gene Manipulation

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Cell culture: Pooled human umbilical vein endothelial cells (HUVECs) were purchased from Lonza (#CC- 5 2519) and cultured in endothelial basal medium (EBM, Lonza) supplemented with hydrocortisone (1μg/ml), bovine brain extract (12 μg/ml), gentamicin (50 μg/ml), human recombinant epidermal growth factor (10 ng/ml), and 10% fetal bovine serum (FBS, Life Technologies). HUVECs were tested negative for mycoplasma and cultured until the fourth passage. Cells were maintained at 37 °C in a humidified atmosphere with 5% CO2.
siRNAs directed against KRIT1 (s2510, Invitrogen) were used for the knockdown experiments at a concentration of 10nM. For overexpression studies, HUVECs were infected with a control lentivirus encoding either GFP, or mutant human PIK3CA with the H1047R mutation (constructed from pBabe puro HA PIK3CA H1047R from Jean Zhao (Addgene plasmid # 12524). Cells were harvested 48 hours post-infection or post-transfection and total RNA was isolated using TRIzol Reagent (Life Technologies). cDNA was generated from 500ng total RNA using the SuperScript IV VILO Master Mix (Thermo Fisher, 11756050) and real time qPCR was performed using Power SYBR Green PCR Master Mix (Applied Biosciences). Protein was harvested with RIPA buffer with complete protease inhibitor cocktail (Roche) and PhosSTOP phosphatase inhibitor cocktail (Roche).

Ren A.A., Snellings D.A., Su S.Y., Hong C.C., Castro M., Tang A.T., Detter M.R., Hobson N., Girard R., Romanos S., Lightle R., Moore T., Shenkar R., Benavides C., Beaman M.M., Mueller-Fielitz H., Chen M., Mericko P., Yang J., Sung D.C., Lawton M.T., Ruppert M., Schwaninger M., Körbelin J., Potente M., Awad I.A., Marchuk D.A, & Kahn M.L. (2021). PIK3CA and CCM mutations fuel cavernomas through a cancer-like mechanism. Nature, 594(7862), 271-276.

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