Heparin

Manufactured by Lonza

Sourced in Switzerland, United States

Heparin is a laboratory equipment product manufactured by Lonza. It is an anticoagulant substance derived from animal tissues, primarily used for analysis and research purposes.

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Lab products found in correlation

Ascorbic acid, by Lonza (31 mentions) Hydrocortisone, by Lonza (28 mentions) Vascular endothelial growth factor (vegf), by Lonza (23 mentions) Human epidermal growth factor (hegf), by Lonza (19 mentions) Fetal bovine serum (fbs), by Lonza (15 mentions) R3 igf 1, by Lonza (14 mentions) Hfgf b, by Lonza (14 mentions) Ga 1000, by Lonza (12 mentions) Huvecs, by Lonza (10 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions) Amphotericin b, by Lonza (6 mentions) Gentamicin, by Lonza (6 mentions) Ebm 2, by Lonza (5 mentions) Gentamicin amphotericin b, by Lonza (5 mentions) Ebm 2 medium, by Lonza (4 mentions) Egm 2, by Lonza (4 mentions) Penicillin, by Lonza (3 mentions) Streptomycin, by Lonza (3 mentions) Insulin like growth factor 1, by Lonza (3 mentions) R igf 1, by Lonza (3 mentions)

Spelling variants of this product

EGM-2 SingleQuots supplement excluding Heparin (2 citations) Bovine brain extract with heparin (2 citations)

47 protocols using heparin

EGFR-Mutant NSCLC Cell Lines and HUVEC Culture

Cited 1 time

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The NSCLC cell lines A549 (ATCC CCL-185), H460 (ATCC HTB-177), H1650 (ATCC CRL-5883) and H1975 (ATCC CRL-5908) were purchased from American Type Culture Collection (Manassas, VA, USA). H1650 and H1975 cell lines have EGFR mutations (delE746-A750 for H1650; L858R and T790 M for H1975) (14 (link)). Human umbilical vein endothelial cells (HUVECs; H-UV001) were purchased from Bioresource Collection and Research Center (Hsinchu City, Taiwan). Cells were grown in complete growth medium [Dulbecco's modified Eagle's medium (Lonza, Basel, Switzerland) for A549 and H460 cells; RPMI-1640 media (Lonza) for H1650 and H1975 cells] supplemented with 10% fetal calf serum (Thermo Fisher Scientific, Inc., Waltham, MA, USA), 30 ng/ml EGF (Invitrogen; Thermo Fisher Scientific, Inc.), 10 U/ml penicillin and 10 µg/ml streptomycin at 37°C and 5% CO₂. Cells were incubated for 72 h and the supernatant of growth medium was collected for detection of VEGF levels. HUVECs were grown in 90% Medium 199 (Lonza) with 25 U/ml heparin (Lonza) and 30 µg/ml endothelial cell growth supplement (Lonza), adjusted to contain 1.5 g/l sodium bicarbonate, 10 U/ml penicillin and 10 µg/ml streptomycin, as well as 10% fetal calf serum. Gefitinib and G418 were purchased from Sigma Aldrich (EMD Millipore, Billerica, MA, USA).

Hung M.S., Chen I.C., Lin P.Y., Lung J.H., Li Y.C., Lin Y.C., Yang C.T, & Tsai Y.H. (2016). Epidermal growth factor receptor mutation enhances expression of vascular endothelial growth factor in lung cancer. Oncology Letters, 12(6), 4598-4604.

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Isolation and Culture of Endothelial Progenitor Cells

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Peripheral blood mononuclear cells from healthy donors were isolated by density gradient centrifugation (Lymphoprep, Axis-Shield PoC, Oslo, Norway), washed with PBS (GIBCO, Invitrogen, Carlsbad, CA) plus 20% FBS, and suspended in endothelial cell basal medium-2 (EBM-2) (Lonza, Allendale, NJ, USA) supplemented with single aliquots of endothelial cell growth medium (EGM-2) (VEGF, human EGF-B, recombinant IGF-1, human EGF, heparin, ascorbic acid, and GA-100, Lonza) and 15% autologous plasma. Mononuclear cells were plated onto fibronectin (Biocoat, BD Biosciences, Franklin Lakes, NJ, USA) in coated, six-well plates at a density of 5 × 10⁶ cells/well. We then incubated the fibronectin-coated plates at 37°C in a 5% CO₂ atmosphere; 4 days later, we removed the cells in suspension, and the fraction of attached cells was cultivated with EBM-2 supplemented with 15% autologous plasma. The medium was recharged every 2 days for 3–4 weeks. After this period, EPCs could be visualized with an optical microscope (OPTIKA Microscopes, Italy) in the form of colonies (colony forming units, CFUs). The EPC phenotype (CD34+CD133+VEGFR2+) was verified with a cellular purity of >90%.

Luna C., Carmona A., Alique M., Carracedo J, & Ramirez R. (2015). TNFα-Damaged-HUVECs Microparticles Modify Endothelial Progenitor Cell Functional Activity. Frontiers in Physiology, 6, 395.

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PLGA Nanoparticles for HUVEC Targeting

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RESOMER^® RG 503 H Poly (D,L-lactide-co-glycolide) acid terminated with 50:50 monomer ratio with a molecular weight of 24,000–38,000 Da (Cat. No. 719870), N-(3-Dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC; Cat. No. E7750), 2-(N-morpholino)ethanesulfonic acid (MES; Cat No. M3671), N-Hydroxysuccinimide (NHS; Cat. No. 130672), and polyvinyl alcohol (PVA; molecular weight 31,000–50,000 Da; 87–89% hydrolyzed; Cat. No. 363073) were obtained from Sigma-Aldrich (St. Louis, MO). Dichloromethane (DCM) and Nile red (99%, ACROS Organics™) were purchased from Fisher Scientific (Waltham, MA). DiR dye (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide; Cat. No.60017) was purchased from Biotium (Fremont, CA). Human umbilical vein endothelial cells (HUVEC) (Cat. No. C2517A Lot No. 0000439577), EGM-2 (endothelial growth medium) (Cat. No.CC-3162), and heparin were purchased from Lonza (Basel, Switzerland). Hoechst stain (Hoechst 33342) and CellMask™ Deep Red stain (Cat. No. C10046) were procured from Life Technologies (now part of Thermo Fisher Scientific, Waltham, MA). YSA (sequence-YSAYPDSVPMMS; purity > 98 % theoretical pI/Mw: 3.80/1347.52) was purchased from GenScript (Piscataway, NJ). Bleomycin was purchased from Hospira, Inc. (Lake Forest, IL).

Patil M.A., Upadhyay A.K., Hernandez-Lagunas L., Good R., Carpenter T.C., Sucharov C.C., Nozik-Grayck E, & Kompella U.B. (2018). Targeted Delivery of YSA-functionalized and Non-functionalized Polymeric Nanoparticles to Injured Pulmonary Vasculature. Artificial cells, nanomedicine, and biotechnology, 46(SUP3), S1059-S1066.

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miRNA Inhibition and H2O2 Stress in HAECs

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Human aortic endothelial cells (HAECs) of passages 3 to 7 donated from 2 males (aged 49 and 50; Lonza; Walkersville, MD) were cultured in EGM2 media supplemented with 10% (v/v) fetal bovine serum (FBS) and supplements (Hydrocortisone, hFGF-B, VEGF, R3-IGF-1, Ascorbic Acid, hEGF, GA-1000 and Heparin, all reagents from Lonza, USA). Cells were grown in a humidified atmosphere at 5% CO₂ and 37°C. miRNA inhibitors and negative controls (50 µmol/L; 100 pmol/well in six-well plate, Life Technologies Corporation, Grand Island, NY 14072, USA) transfection into HAECs was performed using Lipofectamine RNAiMAX (Thermo Fisher Scientific) according to the manufacturers’ instructions. Transfection was performed 48 h prior to being stimulated with 100 μmol/L H₂O₂ for 24 h in HAECs. Then cells were harvested for subsequent analyses of has-miR-192 expression, and DHFR mRNA and protein expression.

Huang K., Narumi T., Zhang Y., Li Q., Murugesan P., Wu Y., Liu N.M, & Cai H. (2021). Targeting MicroRNA-192–5p, a Downstream Effector of NOXs, Reverses Endothelial Dihydrofolate Reductase Deficiency to Attenuate Abdominal Aortic Aneurysm Formation. Hypertension (Dallas, Tex. : 1979), 78(2), 282-293.

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Culturing Primary Endothelial Cells

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HUVECs were obtained from the National Infrastructure of Cell Line Resource (Beijing, China). NhECs were purchased from Cell Systems (Seattle, WA, USA). ECs were maintained in EBM-2 medium (Lonza, Switzerland) containing 2% foetal bovine serum (FBS), 0.1% hydrocortisone, 0.1% R3IGF, 0.4% hFGF-b, 0.1% VEGF, 0.1% ascorbic acid, 0.1% GA-1000, 0.1% h EGF, 0.1% heparin, 100 U/mL penicillin, and 100 mg/mL streptomycin (Lonza, Switzerland).

Sui T., Qiu B., Qu J., Wang Y., Ran K., Han W, & Peng X. (2021). Gambogic amide inhibits angiogenesis by suppressing VEGF/VEGFR2 in endothelial cells in a TrkA-independent manner. Pharmaceutical Biology, 59(1), 1564-1573.

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Culturing HUVEC and SC Cell Lines

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HUVEC cell line was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) (ATCC® PCS-100-010™) and was maintained in endothelial growth medium (EGM-2) medium contains EBM-2 medium (serum free, growth-factor free), supplemented with 2% fetal bovine serum (FBS), human fibroblast growth factor-B (hFGF-B), human epidermal growth factor (hEGF), human vascular endothelial cell growth factor (hVEGF), long R insulin-like growth factor-1 (R3-IGF-1), ascorbic acid, hydrocortisone, and heparin (Lonza). Cells cultured until passage 10. SC cells (human monocytes/macrophages) were purchased from ATCC (ATCC^® CRL-9855™) and cultured in Iscove’s modified Dulbecco’s medium (Thermo Fisher Scientific, Waltham, MA, USA) with 4 mM l-glutamine and supplemented with 0.05 mM 2-mercaptoethanol, 0.1 mM hypoxanthine and 0.016 mM thymidine and 10% FBS (Euroclone). Cells were stained at 37 °C in 5% CO₂—95% air humidified atmosphere.

Belvedere R., Bizzarro V., Parente L., Petrella F, & Petrella A. (2017). The Pharmaceutical Device Prisma® Skin Promotes in Vitro Angiogenesis through Endothelial to Mesenchymal Transition during Skin Wound Healing. International Journal of Molecular Sciences, 18(8), 1614.

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Cell Culture Protocols for Skin Tissue Research

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We used HaCaT cells, HS27 fibroblasts (FBS), and HUVECs as substitutes for the epithelial cells, FBS and blood vessel endothelial cells present in skin tissue54 (link). The human epidermal keratinocytes (HaCaT, ATCC^®, USA) and human skin Fb HS27 cells (CRL-1634, ATCC^®, USA) were cultured in high-glucose DMEM (WELGENE, South Korea) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY) and 1% penicillin/streptomycin solution (Gibco, Grand Island, NY). The HUVECs (PCS-100-010^™, ATCC^®, USA) were cultured in EGM-2 supplemented with ascorbic acid, vascular endothelial growth factor, 20% FBS, recombinant human Fb growth factor, hydrocortisone, insulin-like growth factor-1, a recombinant analog of human insulin-like growth factor-1, gentamicin–amphotericin, and heparin (Lonza, Walkersville, MD, USA). All experiments used cells from passages 3–10.

Wufuer M., Lee G., Hur W., Jeon B., Kim B.J., Choi T.H, & Lee S. (2016). Skin-on-a-chip model simulating inflammation, edema and drug-based treatment. Scientific Reports, 6, 37471.

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Isolation and Culture of Placental hPSCs and Infant Foreskin HDFs

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hPSCs isolated from donated human placenta were obtained from the Manufacturing Development Center at Wake Forest Institute for Regenerative Medicine (IRB# 00056941) (Zhang et al., 2016 (link); Yim et al., 2019 (link)). hPSCs (passage 6–7) were cultured in hPSC growth medium (hPSC-GM) containing 65% of Alpha Minimum Essential Medium (ά-MEM; Gibco, Amarillo, TX), 17% Amniomax C100 Basal Medium (Gibco), 15% Fetal Bovine Serum (FBS; Gibco), 2% Aminomax Supplement (Gibco), 1% Glutamax (Gibco), and 2.5 μg/ml Gentamicin (Gibco). HDFs isolated from donated infant foreskin were obtained from the Wake Forest Institute for Regenerative Medicine Skin Team (IRB# 00007586) (Jorgensen et al., 2020 (link)). HDFs were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Cytiva; Logan, UT, United States) containing 10% FBS and 1% Penicillin/Streptomycin (P/S; Fischer Scientific, Waltham, MA) (HDF-GM) (passage 5–7) (Jorgensen et al., 2020 (link)). The medium was changed every 2–3 days. Human umbilical vein endothelial cell (HUVECs) (Lonza, Basel, Switzerland) were cultured in HUVEC growth medium (HUVEC-GM) containing Endothelial Cell Basal Medium-2 (EBM-2; Lonza) and EBM-2 SingleQuots (EBM-2-SQ; 2% Fetal Bovine Serum, 0.04% Hydrocortisone, 0.4% hFGF-B, 0.1% VEGF, 0.1% R3-IGF-1, 0.1% Ascorbic Acid, 0.1% hEGF, 0.1% GA-1000, 0.1% Heparin; Lonza) (passage 3). The medium was changed every 2–3 days.

Kim J.H., Green D.S., Ju Y.M., Harrison M., Vaughan J.W., Atala A., Lee S.J., Jackson J.D., Nykiforuk C, & Yoo J.J. (2022). Identification and characterization of stem cell secretome-based recombinant proteins for wound healing applications. Frontiers in Bioengineering and Biotechnology, 10, 954682.

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HUVEC Culture Protocol for Endothelial Cell Research

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Endothelial cells from human umbilical vein (HUVEC) (Lonza, Milan, Italy) were used between passages 2 and 5 and were cultured in endothelial basal medium (EBM2) (Lonza, Milan, Italy) supplemented with 2% FCS, 0.4% hFGF-B, 0.1% VEGF, 0.1% R3-IGF-1, 0.1% ascorbic acid, 0.1% hEGF, 0.1% heparin and 0.04% hydrocortisone (Lonza, Milan, Italy) and with 1% penicillin/streptomycin (Sigma-Aldrich, Milan, Italy), and kept at 37 °C in a 5% CO₂ atmosphere with 95% RH.

Saporito F., Sandri G., Bonferoni M.C., Rossi S., Malavasi L., Del Fante C., Vigani B., Black L, & Ferrari F. (2018). Electrospun Gelatin–Chondroitin Sulfate Scaffolds Loaded with Platelet Lysate Promote Immature Cardiomyocyte Proliferation. Polymers, 10(2), 208.

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Quantifying Neurite Outgrowth and Angiogenesis

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For the quantification of neurite outgrowth, the human neuroblastoma cell line (TGW) was starved overnight and then stimulated with CCR3A for 24 h. The mean neurite length was measured under the inverted microscope using ImageJ software (version 1.52, imagej.nih.gov). The same experiment was performed with 50 ng/mL Brain-derived neurotrophic factor (BDNF) (Peprotech, UK) as a positive control. To evaluate the angiogenic effect of CCR3A on HUVEC, HUVEC were seeded on Matrigel (BD Biosciences, San Jose, USA) in DMEM containing 2% FBS, 5 µg/mL heparin (Lonza), 5 µg/mL ascorbic acid (Lonza), 5 µg/mL hydrocortisone (Lonza) supplemented with or without CCR3A. DMEM containing only 2% FBS was used as a negative control and G-CSF (100 ng/mL) was used as a positive control. The mean length of networks of cords and tube-like structures was measured 5 h after cultivation under an inverted microscope (Leica, 6000B-4, Leica Microsystems GmbH, Wetzlar, Germany) using ImageJ software (version 1.52, imagej.nih.gov).

Zayed M., Iohara K., Watanabe H, & Nakashima M. (2020). CCR3 antagonist protects against induced cellular senescence and promotes rejuvenation in periodontal ligament cells for stimulating pulp regeneration in the aged dog. Scientific Reports, 10, 8631.

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