HaCaT-ras II-4 Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
Telephone: +496221405780
Price range: €€€

€800.00*

Prices excl. taxes plus shipping costs

Product number: 300495

Safety data sheet

Product sheet

Third Party Agreement

Description	HaCaT-ras II-4 cells are a remarkable and extensively studied cellular model in biological science. These cells are derived from spontaneously immortalized human skin keratinocytes, known as HaCaT cells, which were modified through transfection with the c-Ha-ras (EJ) oncogene. The selection of these cells was based on their resistance to G418, a selective antibiotic, as described in the comprehensive study conducted by Boukamp et al. in 1990. One notable characteristic of HaCaT-ras II-4 cells is their tumorigenic nature. When these clonal cells are injected into Balb/c-nu/nu mice, they exhibit a fascinating behaviour by forming highly differentiated and locally invasive squamous cell carcinomas. This unique property allows researchers to explore tumour development and progression mechanisms within a controlled experimental environment. HaCaT-ras II-4 cells are predominantly derived from the Caucasian population, ensuring relevance to a specific ethnic group in scientific investigations. Their origin and characteristics make them an invaluable resource for researchers interested in studying various skin biology and differentiation aspects. These cells possess a partially to fully differentiated phenotype under typical culture conditions. This phenotype is attributed to the abundant presence of calcium in both traditional media and fetal bovine serum, which provides an ideal environment for the cells to exhibit characteristics resembling those of mature skin cells. This feature allows researchers to investigate the intricate processes involved in skin development, wound healing, and epidermal differentiation. With their tumorigenic nature and the ability to replicate skin biology in vitro, HaCaT-ras II-4 cells offer a unique opportunity to explore the molecular pathways associated with skin cancer and other skin-related disorders. By utilizing this exceptional cellular model, researchers can gain deeper insights into the underlying mechanisms of tumorigenesis, invasive potential, and therapeutic interventions. HaCaT-ras II-4 cells are a vital tool for biological science research, specifically in skin biology and differentiation studies. Their origin from spontaneously immortalized human skin keratinocytes, modification with the c-Ha-ras (EJ) oncogene, and subsequent tumorigenic behaviour in mice make them invaluable for investigating skin-related diseases and therapeutic approaches. By harnessing the unique characteristics of HaCaT-ras II-4 cells, researchers can unlock a deeper understanding of skin biology and contribute to advancing medical knowledge and treatment options for various skin disorders.
Organism	Human
Tissue	Skin
Synonyms	HaCaT-ras clone II-4, HaCaT II-4, II-4

Age	62 years
Gender	Male
Ethnicity	Caucasian
Cell type	Keratinocyte
Growth properties	Adherent

Citation	HaCaT-ras II-4 (Cytion catalog number 300495)
Biosafety level	1
NCBI_TaxID	9606
CellosaurusAccession	CVCL_3868

Protein expression	P53 (+), CEA (+),
Tumorigenic	Formation of highly differentiated, locally invasive squamous cell carcinoma in Balb/c-nu/nu mice.
Karyotype	Aneuploid (hypotetraploid)

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	Discard Old Medium: Remove the old medium from the flasks. Wash Cells: Add 3-5 ml of PBS (without calcium and magnesium) to T25 flasks, or 5-10 ml to T75 flasks, to wash the adherent cells. Add EDTA Solution: Cover the cell layer completely with a freshly prepared 0.05% EDTA solution-use 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Incubation: Incubate the flasks at 37 degrees Celsius for 10 minutes. Add Trypsin/EDTA Solution: Following the incubation, add a freshly prepared trypsin/EDTA solution (0.05% trypsin, 0.025% EDTA) to the flasks, ensuring the cells are fully covered-use 1 ml for T25 flasks and 2.5 ml for T75 flasks. Monitor Detachment: Observe the cells, which should detach within 1-2 minutes. Neutralize Trypsin: Add FBS-containing cell culture medium to stop the trypsin activity. Transfer Cells: Dispense the cell suspension into new flasks pre-filled with fresh culture medium.
Seeding density	1 x 10⁴ cells/cm²
Fluid renewal	2 times per week
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.
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.

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.

Required products

Required products

DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate

DMEM (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*

Accutase

Variants: 100 ml

Accutase 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*

PBS

Phosphate-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*

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.

HaCaT-ras II-4 Cells

General information

Characteristics

Regulatory Data

Biomolecular Data

Handling

Quality control / Genetic profile / HLA

Third Party Agreement