HaCaT-ras A5 Cells
€800.00*
Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 106 cells for adherent lines or 5 × 106 cells for suspension lines (refer to the batch CoA for details).
General information
| Description | HaCaT-ras A5 cells are a spontaneously immortalized, non-tumorigenic human skin keratinocyte cell line, instrumental in the study of tumour microenvironment interactions and the progression of skin carcinoma. Originating from a 62-year-old Caucasian male, these cells have undergone clonal selection and mutagenesis, which, coupled with autocrine growth factor regulation, enable the formation of slow-growing, highly differentiated benign cystic tumours in Balb/c-nu/nu mice. This makes them a valuable model for investigating the cellular dynamics and molecular mechanisms of tumour progression in vivo. The HaCaT-ras A5 cells are particularly useful for elucidating the complex interactions between tumour cells and surrounding stromal cells, including fibroblasts, immune cells, and endothelial cells. These interactions are mediated by the secretion of various signalling molecules such as growth factors, cytokines, and proteases, among which interleukin-6 (IL-6) plays a pivotal role. IL-6 is known to become dysregulated in many cancer types, primarily through overexpression or persistent activation of the STAT3 transcription factor. Research has shown that IL-6 stimulation of HaCaT-ras A5 cells significantly increases their proliferation via the JAK/STAT signalling pathway, while fibroblasts remain unaffected due to a more potent inhibition by SOCS3, a negative regulator of this pathway. This differential response has been captured in a mathematical model describing the dynamics of STAT3 and SOCS3, providing a deeper understanding of cell-specific signalling cascades. Furthermore, IL-6 not only directly affects HaCaT-ras A5 cell proliferation but also indirectly influences the cellular environment through the activation of a network of growth factors such as HGF, KGF, VEGF, and IL-8. Gene expression analysis involving over 16,000 genes revealed that IL-6 stimulation upregulates 19 genes related to the interferon signal pathway in both HaCaT-ras A5 cells and fibroblasts, which correlates with the observed growth inhibition in fibroblasts. The discovery of the crucial role of SerpinB4 in the proliferation of HaCaT-ras A5 cells, confirmed through siRNA knockdown experiments, underscores the intricate regulation by IL-6 in both tumour and stromal cells. This comprehensive understanding of IL-6’s roles enhances the potential for developing targeted therapeutic strategies aimed at modulating IL-6 signalling pathways in the tumour microenvironment. Overall, HaCaT-ras A5 cells offer a robust model for exploring the complex interplay within the tumour microenvironment, paving the way for novel approaches in cancer research and therapy development. |
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| Organism | Human |
| Tissue | Skin |
| Synonyms | HaCaT-ras clone A-5, HaCaT A-5, A-5, A5 |
Characteristics
| Age | 62 years |
|---|---|
| Gender | Male |
| Ethnicity | Caucasian |
| Cell type | Keratinocyte |
| Growth properties | Adherent |
Regulatory Data
| Citation | HaCaT-ras A5 (Cytion catalog number 300494) |
|---|---|
| Biosafety level | 1 |
| NCBI_TaxID | 9606 |
| CellosaurusAccession | CVCL_xK16 |
| GMO Status | GMO-S1: This HaCaT-ras A5 line contains a plasmid-borne c-Ha-ras oncogene construct for epithelial transformation research. This classification applies only within Germany and may differ elsewhere. |
Biomolecular Data
| Protein expression | P53 (+), CEA (+), |
|---|---|
| Tumorigenic | Formation of benign tumors in Balb/c-nu/nu mice. |
| Karyotype | Aneuploid (hypotetraploid) |
Handling
| Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
|---|---|
| Supplements | Supplement the medium with 10% FBS |
| Dissociation Reagent | The 1:1 mixture of EDTA (stock. 0.05%) and trypsin (stock: 0.1%) must be prepared each time ahead of detaching the cells using PBS without Ca2+ and Mg2+ to provide a physiologic osmolarity. Ready-to-use mixtures of trypsin/EDTA are not recommended, as this may result in cell clumps. As an alternative, TrypLETM Express (Life Technologies) instead of trypsin/EDTA can be used. The protocol of the manufacturer should be followed. |
| Subculturing |
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| Seeding density | 1 x 104 cells/cm2 |
| Fluid renewal | 2 times per week |
| Freeze medium | As a cryopreservation medium, we 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 |
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| Incubation Atmosphere | 37°C, 5% CO2, humidified atmosphere. |
| Shipping Conditions | Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage. |
| Storage Conditions | For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen. |
Quality Control & Molecular Analysis
| 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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Third Party Agreement
Please note that this cell line is not available under a standard Cytion MTA, as it requires a third-party agreement and/or is subject to negotiation with the original licensor.-
Required products
Required products
Freeze Medium CM-1 - 50 mlCryopreservation Media Variants: 50 mlCytion’s Freeze Medium CM-1 is a state-of-the-art cryopreservation medium designed to ensure the highest level of cell viability and functionality post-thaw. This versatile medium is suitable for a broad spectrum of cell types, including both human and animal cells, making it an essential tool for diverse research applications. Formulated with a meticulously balanced combination of cryoprotectants and essential nutrients, Freeze Medium CM-1 minimizes ice crystal formation and cellular stress during the freezing process, thus preserving cellular integrity.
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.€59.00*DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvateDMEM (Dulbecco's Modified Eagle Medium) is a highly versatile and widely utilized basal medium designed to support the growth of a diverse range of mammalian cells in biological research. It serves as an ideal medium for culturing primary fibroblasts, neurons, glial cells, HUVECs, smooth muscle cells, as well as popular cell lines like HeLa, 293, Cos-7, and PC-12.
What sets DMEM apart from other media is its unique composition. It contains an impressive fourfold increase in amino acid and vitamin concentration compared to the original Eagle's Minimal Essential Medium. Initially developed with low glucose (1 g/L) and sodium pyruvate, DMEM is frequently employed with higher glucose levels, either with or without sodium pyruvate. Notably, DMEM does not contain proteins, lipids, or growth factors, necessitating supplementation. To achieve optimal growth, a common approach is to supplement DMEM with 10% Fetal Bovine Serum (FBS). Additionally, DMEM employs a sodium bicarbonate buffer system, requiring a 5-10% CO2 environment to maintain a physiological pH.
Dulbecco's Modified Eagle Medium (DMEM) is highly regarded among defined media for cell and tissue culture, catering to the growth needs of various adherent cell phenotypes. It surpasses the original Eagle's Medium, developed in the 1950s for cultivating chicken cells, through the enhanced supplementary formulation known as Dulbecco's modification. This modification significantly elevates the content of select amino acids and vitamins up to fourfold compared to the original medium.
In the field of cell culture, DMEM plays a vital role as a growth medium for different cell types, including primary cells, stem cells, and transformed cells. Researchers also employ the modified version of DMEM for a wide array of research applications, such as drug discovery, tissue engineering, and the study of cell signaling pathways.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +8°C, protected from light.
Once opened, store at 4°C and use within 6–8 weeks.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Amino Acids
Glycine
30.00
L-Arginine HCl
84.00
L-Cystine 2 HCl
62.57
L-Glutamine
584.00
L-Histidine HCl H2O
42.00
L-Isoleucine
105.00
L-Leucine
105.00
L-Lysine HCl
146.00
L-Methionine
30.00
L-Phenylalanine
66.00
L-Serine
42.00
L-Threonine
95.00
L-Tryptophan
16.00
L-Tyrosine 2 Na 2H2O
103.79
L-Valine
94.00
Vitamins
Choline chloride
4.00
D-Calcium Pantothenate
4.00
Folic Acid
4.00
myo-Inositol
7.20
Nicotinamide
4.00
Pyridoxal HCl
4.00
Riboflavin
0.40
Thiamine HCl
4.00
Inorganic Salts
CaCl2 2 H2O
265.00
Fe(NO3)3 9 H2O
0.10
KCl
400.00
MgSO4 7H2O
200.10
NaCl
6400.00
NaHCO3
3700.00
NaH2PO4 2H2O
141.73
Other Components
D-Glucose
4500.00
Phenol Red Sodium Salt
15.90
Sodium Pyruvate
110.00€25.00*AccutaseVariants: 100 mlAccutase Cell Dissociation Reagent
- 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.€75.00*Antibiotic/Antimycotic Solution (100x)Product Overview
Volume: 100 ml
Storage: ≤-15°C
Sterility: Sterile-filtered
Antibiotic/Antimycotic Solution (100x) is a sterile, ready-to-use concentrate designed to reduce microbial contamination risks in cell culture and related laboratory applications. This 100x solution contains a well-established combination of penicillin, streptomycin, and amphotericin B—providing broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts, and filamentous fungi. The formulation is suitable for use in eukaryotic cell cultures, bacterial media, and other contamination-sensitive systems, supporting clean and consistent lab operations.
Application and Benefits Optimized for routine research protocols, this solution is widely used to maintain aseptic conditions in cell culture workflows. It offers reliable performance in contamination-sensitive environments, helping researchers reduce the risk of microbial overgrowth without compromising cell health or experimental reproducibility. The sterile-filtered formulation eliminates the need for additional solubilization steps, supporting streamlined media preparation and reducing variability in daily lab procedures.
Usage and Compatibility To achieve standard working concentrations, dilute the solution 1:100 into your complete culture medium. The product is compatible with a broad range of mammalian cell lines and basal media. With consistent stock availability, researchers benefit from dependable supply continuity and simplified logistics planning. The solution should be stored at ≤ –15 °C and protected from repeated freeze-thaw cycles to maintain stability. For research use only. Not for use in diagnostic or therapeutic procedures. Not for use in humans or animals.€45.00*PBSPhosphate-Buffered Saline (PBS) Solution
Phosphate-buffered saline (PBS) is a widely used buffer solution in biological and chemical research. It plays a crucial role in maintaining the pH balance and osmolarity during various experimental procedures, including tissue processing and cell culture. Our PBS solution is meticulously formulated with high-purity ingredients to ensure stability and reliability in every experiment. The osmolarity and ion concentrations of our PBS closely mimic 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:
8000 mg/L Sodium chloride (NaCl)
200 mg/L Potassium chloride (KCl)
1150 mg/L Sodium phosphate dibasic anhydrous (Na2HPO4)
200 mg/L Potassium phosphate monobasic anhydrous (KH2PO4)
This composition ensures an optimal pH and ionic balance, suitable for a wide range of biological applications.
Applications of Our PBS Solution
Our PBS solution is ideal for various applications in biological research. Its isotonic and non-toxic properties make it suitable for substance dilution and cell container rinsing. PBS solutions containing EDTA are effective for disengaging attached and clumped cells. However, divalent metals such as zinc should not be added to PBS, as this can cause precipitation. In such cases, Good's buffers are recommended. Additionally, our PBS solution is an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, including SARS-CoV-2.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +25°C, protected from light.
Once opened, store at 2°C to 25°C and use within 24 months.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Salts
Potassium chloride
200
Potassium phosphate monobasic anhydrous
200
Sodium chloride
8000
Sodium phosphate dibasic anhydrous
1150€20.00*