M2-10B4 Cells

Categories: Mouse cell lines Description: The M2-10B4 cell line is a clone derived from bone marrow stromal cells from a (C57BL/6J X C3H/HeJ)F1 mouse. These stromal cells are essential components of the bone marrow microenvironment and play a significant role in supporting hematopoiesis. The M2-10B4 cells are particularly valuable for research focused on the interactions between stromal and hematopoietic cells, as they can support both human and murine myelopoiesis in long-term culture. Additionally, these cells can sustain certain murine stromal cell-dependent pre-B cell lines in vitro, making them a versatile tool in hematopoietic research. The M2-10B4 cells express important extracellular matrix components such as laminin and collagen IV, which contribute to their ability to support hematopoietic cells. However, they do not express collagen I or Factor VIII, which distinguishes them from other stromal cell lines. The presence of laminin and collagen IV is critical for the maintenance of the bone marrow microenvironment, influencing cell adhesion, differentiation, and signaling pathways. Researchers often utilize the M2-10B4 cell line in co-culture systems to explore the effects of stromal cells on the behavior of hematopoietic progenitors, particularly in the context of bone marrow physiology and disease models. Given their origin and functional properties, M2-10B4 cells are an essential model for studying the bone marrow niche, especially in relation to hematological disorders such as leukemia. They are also useful in drug screening and the development of therapeutic strategies targeting the bone marrow microenvironment. As with all cell lines, it is important to note that M2-10B4 cells are intended for research use only and are not suitable for therapeutic or in vivo applications. Organism: Mouse Tissue: Bone marrow Synonyms: M210B4 Breed: C57BL/6J x C3H/HeJ Age: Unspecified Gender: Female Morphology: Fibroblast-like Cell Type: Fibroblast Growth Properties: Adherent Citation: M2-10B4 (Cytion catalog number 400428) Biosafety Level: 1 Ncbi_ Taxid: 10090.0 Cellosaurus Accession: CVCL_5794 Products: Laminin, collagen IV (Collagen I(-), Factor VIII(-). Culture Medium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) 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. Seeding Density: 1 x 104 cells/cm2 Fluid Renewal: 2 to 3 times per week Post Thaw Recovery: Viability may be low after thawing. 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: 820700a Required Product 3: 860015.0 Required Product 4: 830100.0