HUVEC, single donor

Categories: HUVEC Description: Human Umbilical Vein Endothelial Cells (HUVECs) are primary cells derived from the endothelial layer of veins in the human umbilical cord. HUVECs are a pivotal model in vascular biology research due to their capacity to closely replicate many aspects of endothelial cell biology in vivo. These cells are extensively utilized to study endothelial functions, including angiogenesis, inflammation, and mechanisms of vascular permeability. HUVECs display several critical endothelial markers, such as von Willebrand factor, CD31, and endothelial nitric oxide synthase (eNOS), which affirm their endothelial origin and functionality. They are also capable of forming tube-like structures when cultured on Matrigel, demonstrating their potential for angiogenesis studies. The ability of HUVECs to respond to cytokines and growth factors makes them an excellent system for exploring cellular responses associated with vascular diseases such as atherosclerosis, hypertension, and thrombosis. Moreover, their reaction to shear stress can be studied in dynamic flow models, providing insights into the effects of blood flow on endothelial behavior. In pharmacological research, HUVECs are commonly employed to evaluate the efficacy and toxicity of vascular-targeting agents. Their straightforward isolation and the relative ease of culturing make them a valuable tool in both academic research and pharmaceutical development. These attributes underline the significance of HUVECs in advancing our understanding of vascular health and disease. Organism: Human Tissue: Umbilical vein Synonyms: Human Umbilical Vein Endothelial Cells Ethnicity: Caucasian Morphology: Endothelial Cell Type: Primary cells Growth Properties: Monolayer, adherent Citation: HUVEC, pooled (Cytion catalog number 300605) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Protein Expression: Cytosplasmic VWF/ Factor VIII > 95% positive by immunofluorescence. Cytoplasmic uptake of Di-I-Ac-LDL > 95% positive by immunofluorescence. Cytoplasmic PECAM1 > 95% positive by immunofluorescence Viruses: Negative for HIV-1, HBV, and HCV Culture Medium: Endothelial Cell Growth Medium (PromoCell article number C-22010) Dissociation Reagent: Accutase Subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. Fluid Renewal: Every 2 to 3 days Freeze Medium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. Thawing And Culturing Cells: Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks, for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes. Sterility: Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 3: 860015.0 Required Product 4: 830100.0