Hep-74.3A Cells

€800.00*

Prices excl. taxes plus shipping costs

Product number: 400208

Safety data sheet

Product sheet

Certificate of Analysis (CoA)

Material Transfer Agreement

Description	The Hep-74.3 hepatoma cell line is derived from a mouse liver tumor, specifically from the C3H/He mouse strain. This cell line is characterized by its hepatocytic origin, confirmed through intermediate filament protein analysis. Hep-74.3 expresses simple keratins K8 and K18, which are typical of normal liver cells, as well as vimentin and keratin K19 to varying degrees. These protein patterns confirm the hepatocytic nature of the cell line and its classification as a hepatoma line. The Hep-74.3 cell line displays a predominantly epithelial morphology, reflecting its origin from hepatocytes. This morphological phenotype is consistent with its protein expression profile. DNA fingerprint analysis of Hep-74.3 did not reveal any major structural abnormalities, indicating a degree of genomic stability. However, some changes in the relative intensities of specific bands were observed with increasing passage numbers, suggesting minor genomic variability over extended culture periods. Despite the absence of detectable p53 mutations in the primary mouse liver tumors, aberrations were found in some hepatoma lines during in vitro propagation. The Hep-74.3 cell line was analyzed for mutations in the p53 and c-Ha-ras genes. The absence of detectable mutations in the p53 gene in this line during early passages suggests a stable genetic background. This cell line serves as a valuable model for studying hepatocellular carcinoma, providing insights into the cellular and molecular mechanisms underlying liver tumorigenesis.
Organism	Mouse
Tissue	Liver
Disease	Hepatocellular carcinoma
Synonyms	Hep-74.3, HEP-74.3a, 74.3A, 74.3a

Breed/Subspecies	C57BL/6J
Age	Adult
Gender	Female
Morphology	Epithelial-like
Growth properties	Adherent

Citation	Hep-74.3A (Cytion catalog number 400208)
Biosafety level	1
NCBI_TaxID	10090
CellosaurusAccession	CVCL_5773

Protein expression	Keratin 8, Keratin 18, Vimentin
Tumorigenic	Yes, in C3H/HE mice
Mutational profile	p53 wt

Culture Medium	Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a)
Supplements	Supplement the medium with 10% FBS
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.
Seeding density	1 x 10⁴ cells/cm²
Fluid renewal	Every 3 to 5 days
Post-Thaw Recovery	After thawing, plate the cells at 5 x 10⁴ cells/cm² and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
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

Ham's F12 Medium, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.176 g/L NaHCO3

Ham's F-12 Medium, also known as Ham's F-12 Nutrient Mix, is a widely used basal medium designed specifically for cell culture. It has been extensively employed for serum-free, single-cell plating of Chinese Hamster Ovary (CHO) cells, lung cells, and mouse L cells. Additionally, Ham’s F-12 is the medium of choice for a Clonal Toxicity Assay (CTA).
One of its notable advantages is the ability to support cell growth without the need for serum supplementation. This eliminates potential interference caused by serum components, ensuring consistent and reliable experimental results. By providing a serum-free culture environment, Ham's F-12 Medium offers researchers greater control over their investigations.
Another key feature of Ham's F-12 Medium is its suitability for single-cell plating. This makes it an excellent choice for a variety of cell lines, including CHO cells, lung cells, and mouse L cells. The medium's optimized nutrient composition facilitates efficient attachment and growth of individual cells, enabling the establishment of homogeneous cell cultures with improved reproducibility.
Moreover, Ham's F-12 Medium has gained recognition as the preferred medium for the Clonal Toxicity Assay (CTA). This assay plays a critical role in assessing the cytotoxic effects of substances on cells. By utilizing Ham's F-12 Medium in the CTA, researchers can accurately evaluate the impact of various compounds or treatments on individual cells, providing valuable insights into toxicological profiles.
Quality control

pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.

Maintenance

Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.

Composition

Components
mg/L

Inorganic Salts
Calcium chloride x 2H2O
44,00

Copper(II) sulfate x 5H2O
0,00

Iron (II) sulfate x 7H2O
0,83

Magnesium chloride x 6H2O
122,00

Potassium chloride
223,65

Sodium chloride
7599,00

di-Sodium hydrogen phosphateanhydrous
142,04

Zinc sulfate x 7H2O
0,86

Other Components
D(+)-Glucose anhydrous
1801,60

Hypoxanthine
4,08

Linoleic acid
0,08

DL-α-Lipoic acid
0,21

Phenol red
1,20

Putrescine x 2HCl
0,16

Sodium pyruvate
110,00

Thymidine
0,73

NaHCO3
1176,00

Amino Acids
L-Alanine
8,91

L-Arginine x HCl
210,70

L-Asparagine x H2O
15,01

L-Aspartic acid
13,31

L-Cysteine x HCl x H2O
35,12

L-Alanyl-L-Glutamine
217,30

L-Glutamic acid
14,71

Glycine
7,51

L-Histidine x HCl x H2O
20,96

L-Isoleucine
3,94

L-Leucine
13,12

L-Lysine x HCl
36,54

L-Methionine
4,48

L-Phenylalanine
4,96

L-Proline
34,53

L-Serine
10,51

L-Threonine
11,91

L-Tryptophan
2,04

L-Tyrosine
5,44

L-Valine
11,71

Vitamins
D(+)-Biotin
0,01

D-Calcium pantothenate
0,24

Choline chloride
13,96

Folic acid
1,32

myo-Inositol
18,02

Nicotinamide
0,04

Pyridoxine x HCl
0,06

Riboflavin
0,04

Thiamine x HCl
0,34

Vitamin B12
1,36

€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*

Lot Number	Certificate Type	Date	Catalog Number
400208-313	Certificate of Analysis	23. May. 2025	400208

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.

Hep-74.3A Cells

General information

Characteristics

Regulatory Data

Biomolecular Data

Handling

Quality control / Genetic profile / HLA

Certificate of Analysis (CoA)

Material Transfer Agreement