MC3T3-E1 Subclone 24 Cells

Categories: Mouse cell lines Description: MC3T3-E1 Subclone 24 cells expressly represent a preosteoblast cell type, which plays a crucial role in bone formation. Morphologically, they exhibit a fibroblast-like appearance, characterized by their elongated shape and spindle-like structures. This particular subclone is derived from the calvaria tissue, a skull region contributing to bone formation. One of the critical applications of MC3T3-E1 Subclone 24 Cells lies in 3D cell culture, where researchers can study the behavior and interactions of these cells within a three-dimensional environment. This method offers a more physiologically relevant model than traditional two-dimensional cell cultures, allowing for a better understanding of the intricate processes involved in bone formation. While these cells possess numerous advantages, it's important to note their specific characteristics. MC3T3-E1 Subclone 24 Cells have been observed to exhibit poor osteoblast differentiation when exposed to ascorbic acid, a key component for promoting bone cell growth. Furthermore, they do not form a mineralized extracellular matrix, a crucial step in creating bone tissue. The doubling time of MC3T3-E1 Subclone 24 Cells is approximately 90.5 hours. Organism: Mouse Tissue: Bone Breed: C57BL/6 Age: 1 day Gender: Unspecified Morphology: Fibroblast Cell Type: Osteoblast Growth Properties: Adherent Citation: MC3T3-E1 Subclone 24 (Cytion catalog number 305186) Biosafety Level: 1 Ncbi_ Taxid: 10090.0 Cellosaurus Accession: CVCL_5438 Receptors Expressed: parathyroid hormone-related protein (PTHrP) receptor Protein Expression: Collagen, bone sialoprotein (BSP), osteocalcin (OCN), parathyroid hormone (PTH) Tumorigenic: Yes, in immunosuppressed mice Culture Medium: Alpha MEM, w: 2.0 mM stable Glutamine, w: Ribonucleosides, w: Deoxyribonucleosides, w: 1.0 mM Sodium pyruvate, w: 2.2g/L NaHCO3, w/o: Ascorbic acid (GIBCO, Catalog No. A1049001. We do not supply this product, please consider other suppliers. Please let us know if you need further assistance.) 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. 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. 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. 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 3: 860015.0 Required Product 4: 830100.0