MDBK (NBL-1) Cells

Categories: Bovine cell lines Description: MDBK cells, short for Madin-Darby Bovine Kidney cells (also known as NBL-1), are an exceptional biological resource derived from the kidneys of apparently healthy adult Bos taurus, specifically male individuals. These cells grow adherently and possess an epithelial-like morphology. One of the remarkable applications of MDBK cells lies in their ability to facilitate in vitro studies on the expression of Eimeria bovis-derived antigens on the host cell surface membrane. Additionally, MDBK cells have been employed in investigations centred around the ubiquitination and degradation of signal transducer and activator of transcription 1 and 2 (STAT1 and STAT2) by the V proteins of paramyxoviruses, such as simian virus five and human parainfluenza virus type 2. With an average doubling time ranging from 24 to 35 hours, MDBK cells exhibit a moderate proliferation rate. The establishment of the MDBK cell line dates back to February 18, 1957, when S.H. Madin and N.B. Darby successfully derived it from the kidney of a healthy adult steer. Since then, these cells have become a cornerstone in biological research, enabling numerous breakthroughs in various scientific fields. The karyotype analysis of MDBK cells reveals a modal chromosome number of 51, indicating a hypodiploid state. Within the cell population, the hypodiploid condition manifests as a stemline chromosome number of 2n = 60, with a 2S component occurring in approximately 5% of the cells. Moreover, 11-14 marker chromosomes are typically present, comprising a combination of metacentric, submetacentric, and acro-telocentric chromosomes. Notably, the x chromosome appears monosomic, while no HSR chromosomes or DM's (double minutes) are observed. MDBK cells exhibit an array of applications in the realm of biological research. Their utility extends to 3D cell culture, enabling scientists to recreate complex tissue-like structures for advanced studies. Furthermore, MDBK cells are invaluable in high-throughput screening, facilitating the rapid and efficient screening of compounds or agents for various purposes. Additionally, these cells play a crucial role in toxicology studies, essential for evaluating the safety and potential adverse effects of substances on living organisms.Regarding viral susceptibility, MDBK cells demonstrate receptiveness to several pathogens, including Vesicular stomatitis Orsay (Indiana) virus, infectious bovine rhinotracheitis virus, bovine rhinotracheitis virus, bovine parvovirus, bovine adenovirus 2 and 3, bovine viral diarrhoea virus 1, and parainfluenza three virus. This susceptibility to a diverse range of viruses makes MDBK cells invaluable for investigating viral pathogenesis and evaluating antiviral strategies. Organism: Bovine Tissue: Kidney Synonyms: MDBK (NBL-1), NBL-1, Madin-Darby Bovine Kidney, Madin Darby Bovine Kidney Breed: Bos taurus Age: Adult Gender: Male Morphology: Epithelial-like Growth Properties: Monolayer, adherent Citation: MDBK (NBL-1) (Cytion catalog number 600396) Biosafety Level: 1 Ncbi_ Taxid: 9913.0 Cellosaurus Accession: CVCL_0421 Viruses: The line was tested and shown to be free of bovine diarrhoea virus (BVD). Virus Susceptibility: The cells are susceptibled to bovine diarrhea virus, vesicular stomatitis (Indiana strain), infectious bovine rhinotracheitis virus, bovine parvovirus, bovine adenovirus I and III, and parainfluenza virus 3. Virus Resistance: Poliovirus 2 Reverse Transcriptase: negative Products: Keratin 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: 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 days Post Thaw Recovery: Fast 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