OP9 Cells
Product number:
305174
Characterization of the OP9 cell line
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-?1, and TGF-?3, 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. |
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Organism | Mouse |
Tissue | Bone marrow, stroma |
Synonyms | OP-9 |
Characteristics
Age | Embryo |
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Morphology | Fibroblast-like |
Growth properties | Adherent |
Documentation
Citation | OP9 (Cytion catalog number 305174) |
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Biosafety level | 1 |
Genetic makeup
Culturing methods of OP9 stromal cells
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 |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | 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. |
Split ratio | 1:2 to 1:4 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | OP9 cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality assurance for the authentic mesenchymal stem cell line OP9
Sterility | Mycoplasma contamination is rigorously 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. |
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