HUVEC, single donor
General information
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. |
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Organism | Human |
Tissue | Umbilical vein |
Applications | Human Umbilical Vein Endothelial Cells (HUVECs) are widely used in various biomedical research areas because they can quickly proliferate and differentiate into different types of endothelial cells, which line blood vessels. HUVECs have many research and drug discovery applications, including wound healing, angiogenesis, tissue engineering, inflammation, oncology, pharmacology, vascular modeling, and transfection. |
Synonyms | Human Umbilical Vein Endothelial Cells |
Characteristics
Ethnicity | Caucasian |
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Morphology | Endothelial |
Cell type | Primary cells |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | HUVEC, pooled (Cytion catalog number 300605) |
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Biosafety level | 1 |
Expression / Mutation
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 |
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Viruses | Negative for HIV-1, HBV, and HCV |
Handling
Culture Medium | Endothelial Cell Growth Medium (Cytion article number 820731) |
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Passaging solution | 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. |
Split ratio | A ratio of 1:2 to 1:4 is recommended |
Fluid renewal | Every 2 to 3 days |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
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. |
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Benötigte Produkte
Key features of Freeze Medium CM-1 include:
Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-1 for Freezing Cells
To use CM-1 for freezing both adherent and suspension cells, follow these steps:
For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.
🥶 Method
🔍 Description
💡 Steps
❄️
Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.
🧊
Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.
🧬
Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.
Key features of Freeze Medium CM-1 include:
Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-1 for Freezing Cells
To use CM-1 for freezing both adherent and suspension cells, follow these steps:
For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.
🥶 Method
🔍 Description
💡 Steps
❄️
Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.
🧊
Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.
🧬
Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.
Key features of Freeze Medium CM-1 include:
Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-1 for Freezing Cells
To use CM-1 for freezing both adherent and suspension cells, follow these steps:
For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.
🥶 Method
🔍 Description
💡 Steps
❄️
Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.
🧊
Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.
🧬
Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a versatile buffer solution used in many biological and chemical applications, as well as tissue processing. Our PBS solution is formulated with high-quality ingredients to ensure a constant pH during experiments. The osmolarity and ion concentrations of our PBS solution are matched to those of the human body, making it isotonic and non-toxic to most cells.
Composition of our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains 137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 2 mM KH2PO4. We have chosen this composition based on CSHL protocols and Molecular cloning by Sambrook, which are well-established standards in the research community.
Applications of our PBS Solution
Our PBS solution is ideal for a wide range of applications in biological research. Its isotonic and non-toxic properties make it perfect for substance dilution and cell container rinsing. Our PBS solution with EDTA can also be used to disengage attached and clumped cells. However, it is important to note that divalent metals such as zinc cannot be added to PBS as this may result in precipitation. In such cases, Good's buffers are recommended. Moreover, our PBS solution has been shown to be an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, such as SARS-CoV-2.
Storage of our PBS Solution
Our PBS solution can be stored at room temperature, making it easy to use and access.
To sum up
In summary, our PBS solution is an essential component in many biological and chemical experiments. Its isotonic and non-toxic properties make it suitable for numerous applications, from cell culture to viral transport medium. By choosing our high-quality PBS solution, researchers can optimize their experiments and ensure accurate and reliable results.
Composition
Components
mg/L
Inorganic Salts
Potassium chloride
200,00
Potassium dihydrogen phosphate
200,00
Sodium chloride
8,000.00
di-Sodium hydrogen phosphate anhydrous
1,150.00
Definition of Endothelial Cell Growth Media
An in-vitro cell culture system requires a particular solution called Endothelial Cell Growth Media to sustain the endothelial cells necessary for the experiment. Endothelial cells in the body are exposed to an environment mimicked by the medium, including necessary and non-essential amino acids, vitamins, hormones, growth factors, and trace minerals. This nutrient-dense fluid is critical for preserving the endothelium phenotype and function because it stimulates cell proliferation. Globally, researchers and scientists rely on the CLS Endothelial Cell Growth Medium because it is both effective and simple.
Importance of Endothelial Cell Research
Blood artery creation, blood pressure control, and wound healing are just a few physiological processes in which endothelial cells play a key part. To keep the blood vessels balanced, these cells may react to chemical and mechanical stimuli. In order to fully comprehend the causes of hypertension, atherosclerosis, and thrombosis, research into endothelial cells is crucial. Investigating endothelial cells may also provide light on how to treat these diseases medically. Providing a stable and uniform environment for endothelial cell development and proliferation, the CLS Endothelial Cell Growth Medium has shown to be an indispensable tool in this kind of study.
Quality Assurance
High-quality in-vitro cell culture media is essential for obtaining consistent and repeatable outcomes. Quality control testing is performed on every CLS Endothelial Cell Growth Media batch to guarantee its uniformity and effectiveness. The sterility, lack of mycoplasma and bacteria, and pH levels of each batch are examined. To promote healthy cell development and proliferation, the medium's pH is kept at 7.2 +/
- 0.02, and the temperature is maintained between 20 and 25 degrees Celsius. To ensure the culture is clear of any contamination that might impact the accuracy of the findings, the medium is also checked for microbiological pollutants such as fungus, bacteria, and mycoplasma.
Maintenance and Disclaimer
The expiration date of CLS Endothelial Cell Growth Media is six weeks from the production date. The medium should be kept at a temperature of between +2°C and +8°C, away from light, and never frozen or heated over 37°C to preserve its quality. As a corollary, it is crucial to avoid microwaves and other unregulated heat sources that might compromise the product's integrity. A portion of the medium may be taken out of the container and brought up to room temperature if just a little amount will be utilized. CLS's Endothelial Cell Growth Medium is a tried and true method for cultivating endothelial cells because of the stringent quality controls used in its production.
Notably, the CLS Endothelial Cell Growth Medium is not for clinical or diagnostic use; it is designed primarily for in-vitro usage. Obtaining accurate and reliable findings from the medium requires strict adherence to the manufacturer's guidelines and quality control methods.
Applications of the Medium
The CLS Endothelial Cell Growth Medium has proven useful in several biological studies. As this medium may be used to cultivate endothelial cells, it can be used to simulate endothelial function and malfunction in organ systems in vitro. This is especially useful for studying the blood-brain barrier or tissue-engineered blood arteries. This is essential for understanding disease processes and creating effective treatments.
Endothelial cells, especially HUVEC cells, have been extensively employed in angiogenesis research, wound healing research, and cancer research, thus, the medium has also been modified to promote their proliferation. Accurate and repeatable findings depend on the consistency and high quality of the medium used to develop these cells, and the CLS Endothelial Cell Growth Media meets both of these requirements.