LMH Cells

Categories: Avian cell lines Description: LMH cells, derived from a Leghorn male hepatoma, are a versatile cell line widely used in biological research. Tomoyuki Kitagawa established them in 1981 at the Cancer Institute in Tokyo, Japan. These cells have an epithelial phenotype and are particularly useful for studying host-pathogen interactions in the gastrointestinal tract of poultry. LMH cells are adherent and exhibit a dendritic-like morphology. They express glucose-6-phosphatase and weak canalicular ATPase activity. With a triploid karyotype and six marker chromosomes, these cells display distinct genetic characteristics.Notably, LMH cells have been shown to efficiently support duck hepatitis B virus (DHBV) DNA synthesis when transfected with viral constructs. This makes them an invaluable tool for virology research, particularly in the context of poultry-related viral infections.The derivation of LMH cells involved inducing tumorous nodules in the liver of Leghorn chickens through long-term treatment with diethylnitrosamine. These cells have also been chemically transformed, allowing for their immortalization and continuous propagation in culture. In terms of tumorigenicity, LMH cells have the ability to form tumors in athymic nude mice. This characteristic makes them an important model for studying hepatocellular carcinoma. LMH cells express the estrogen receptor and can be induced to express the liver-specific apolipoprotein II (apoII) gene. This indicates their involvement in estrogen signaling pathways and lipid metabolism. To culture LMH cells, it is necessary to precoat tissue culture vessels with collagen. This ensures proper cell adhesion and growth. Organism: Chicken Tissue: Liver Disease: Hepatocellular carcinoma Synonyms: Leghorn Male Hepatoma cell line Breed: Leghorn Age: 16 months Gender: Male Morphology: Epithelial-like, Dendritic like. Growth Properties: Adherent. It may take a couple of days until cells grow in fully adherent colonies. Citation: LMH (Cytion catalog number 601411) Biosafety Level: 1 Ncbi_ Taxid: 9031.0 Cellosaurus Accession: CVCL_2580 Receptors Expressed: Estrogen (low level expression). Tumorigenic: LMH cells form tumors in athymic mice. Products: glucose-6-phosphatase, canalicular ATPase activity (weak) Karyotype: triploid, modal number = 116, six marker chromosomes Culture Medium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a) Supplements: Supplement the medium with 10% FBS and 1% NEAA Dissociation Reagent: Accutase Subculturing: LMH cells attach better to tissue culture vessels which have been precoated with Collagen. Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium. Seeding Density: 1 to 3 x 104 cells/cm2 Fluid Renewal: Every 2 days 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. 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. Certificate Of Analysis: 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. Subject To Material Transfer Agreements: 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. Required Product 1: 820100a Required Product 3: 860015.0 Required Product 4: 830100.0