A427 Cells

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Profit Organisation/Nutzung: CLS_CLEAR.pdf
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ArtNr	CLS-300111
Hersteller	CLS Cell Lines Service
Menge	1 cryovial
Kategorie	Cell Lines & Cell Culture Cell Lines
Typ	Cell line
Zertifikat	For a certificate of analysis, please send an inquiry to info@hoelzel.de.
Specific against	Human (Homo sapiens)
Dry ice	Yes
ECLASS 10.1	42040401
ECLASS 11.0	42040401
UNSPSC	41106509
Alias	A-427,A427N
Lieferbar
Manufacturer - Category	Lung cancer cell lines
Description	A427 cells originate from lung tissue, specifically a carcinoma, exhibit epithelial morphology and grow adherently. A427 cells have a doubling time of approximately 28 hours in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). In ACL-3 medium, the doubling time is slightly extended to 38 hours, while in ACL-3 supplemented with bovine serum albumin (BSA), it reaches 42 hours. These variations in doubling time provide valuable insights into cell behaviour under different experimental conditions. At passage 60, A427 cells display a hypotriploid to hypertriploid karyotype. This means the cells possess abnormal chromosomes, including dicentrics, minutes, and a large subtelocentric marker. Such karyotypic abnormalities are often associated with cancer cells and contribute to the unique characteristics of this cell line. A427 cells exhibit tumorigenic properties, allowing them to form tumours when injected into nude mice. These tumours resemble undifferentiated adenocarcinoma, further emphasizing the relevance of this cell line in studying lung cancer and its progression. With its exceptional features, A427 cells find utility in various applications, particularly in cancer research. Their epithelial morphology and lung origin make them an ideal model for studying lung cancer and related diseases. Additionally, A427 cells are well-suited for 3D cell culture techniques, providing a more physiologically relevant environment to explore the behaviour of lung cancer cells.
Tissue	Lung
Growth properties	Adherent
Disease	Carcinoma
Age	52 years
Gender	Male
Ethnicity	Caucasian
Morphology	Epithelial-like
Biosafety Level	1
Culture Medium	EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c)
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	2 to 3 times per week
Freezing Recovery	After thawing, plate the cells at 4 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.
Amelogenin	X, Y
Tumorigenic	Yes, in nude mice. Forms an undifferentiated tumor suggestive of adenocarcinoma.
Split Ratio	A ratio of 1:3 to 1:5 is recommended
Seeding Density	1 x 10^4 cells/cm^2 will result in a confluent monolayer within 3 days.
Protein Expression	p53 positive
Karyotype	P60) hypotriploid to hypertriploid with abnormalities including dicentrics, minutes and large subtelocentric marker

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Menge: 1 cryovial

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