Hep-74.3A Cells

ArtNr	CLS-400208
Hersteller	CLS Cell Lines Service
Menge	1 cryovial
Kategorie	Cell Lines & Cell Culture Cell Lines
Typ	Cell line
Zertifikat	The certificate of analysis can be requested on the website or via email at info@cytion.com. Please indicate the lot number of your product in the email.
Specific against	Mouse (Murine, Mus musculus)
Dry ice	Yes
ECLASS 10.1	42040401
ECLASS 11.0	42040401
UNSPSC	41106509
Alias	Hep-74.3, HEP-74.3a, 74.3A, 74.3a
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Manufacturer - Category	Mouse cell lines
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.
Tissue	Liver
Growth properties	Adherent
Disease	Hepatocellular carcinoma
Age	Adult
Gender	Female
Breed	C57BL/6J
Morphology	Epithelial-like
Biosafety Level	1
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)
Medium Supplements	Supplement the medium with 10% FBS
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 3 to 5 days
Freezing Recovery	After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
Freeze Medium	CM-1 (Cytion catalog number 800100)
Handling of Cryopreserved Cultures	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.
Disclaimer	Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting.
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.
Tumorigenic	Yes, in C3H/HE mice
Split Ratio	A ratio of 1:4 to 1:8 is recommended
Seeding Density	1 x 10^4 cells/cm^2
Mutational Profile	p53 wt
Protein Expression	Keratin 8, Keratin 18, Vimentin
NOTE	Deutsch: Universitäre Kunden: Für den Erwerb ist ein Material Transfer Agreement oder eine Limited Use Label License auszufüllen. Kommerzielle Kunden: Für den Erwerb ist ein Material Transfer Agreement oder ein Master Supply Agreement auszufüllen. Nach eingegangener Bestellung werden ihnen alle relevanten Dokumente zugeschickt. English: University Customers: A Material Transfer Agreement or Limited Use Label License must be completed for purchase. Commercial Customers: A Material Transfer Agreement or Master Supply Agreement must be completed for purchase. After the order is received, all relevant documents will be sent to you.

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Menge: 1 cryovial

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