OP9 Cells

Categories: Mouse cell lines Description: The OP9 cell line, a stromal cell line derived from the calvariae of op/op mice, has a mutation that leads to a lack of macrophage colony-stimulating factor (M-CSF), which is a critical cytokine involved in the differentiation, survival, and function of various cell types, including macrophages and osteoclasts. OP9 cells have been extensively used in the field of hematopoiesis research as feeder layers in co-culture systems to support the differentiation and expansion of both hematopoietic stem cells (HSCs) and embryonic stem cells (ESCs). These co-culture systems have facilitated the study of hematopoietic differentiation pathways, enabling MSCs to differentiate into adult erythroid cells, erythroblasts, and red blood cells and osteocytes, chondrocytes, myocytes, tenocytes, and adipocytes. The supportive role of OP9 cells in these systems is attributed to their ability to produce a conducive microenvironment rich in cytokines and growth factors essential for stem cell proliferation and lineage-specific differentiation. Furthermore, the OP9 cell line is instrumental in studying the leukocyte reaction and the development of immune cells such as natural killer (NK) cells, demonstrating the OP9 mouse line's utility in immunological research. The secretory factors produced by OP9 cells, including growth factors like bFGF, IGF-1, IL-3, PDGF-BB, TGF-beta1, and TGF-beta3, play a critical role in cell migration and differentiation processes. OP9 cells exhibit a fibroblast-like appearance, characterized by a spindle-shaped, flat morphology. This morphological trait is typical of mesenchymal stromal cells, which are known for their supportive functions in the bone marrow microenvironment. Despite their vast potential, OP9 cells have limitations due to their non-immortalized nature, which confines their use to short-term and small-scale projects, underscoring the need for careful planning and consideration in experimental designs. Organism: Mouse Tissue: Bone marrow, stroma Synonyms: OP-9 Breed: (C57BL/6 x C3H) F2-op/op Age: Embryo Morphology: Fibroblast-like Growth Properties: Adherent Citation: OP9 (Cytion catalog number 305174) Biosafety Level: 1 Ncbi_ Taxid: 10090.0 Cellosaurus Accession: CVCL_4398 Culture Medium: Alpha MEM, w: 2.0 mM stable Glutamine, w/o: Ribonucleosides, w/o: Deoxyribonucleosides, w: 1.0 mM Sodium pyruvate, w: 2.2g/L NaHCO3 Supplements: Supplement the medium with 20% 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. Fluid Renewal: 2 to 3 times per week 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