| Field | Specification |
|---|---|
| Mfr No | |
| Species |
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-β1, and TGF-β3, 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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- cultureMedium: 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
- dissociationReagent: 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.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: 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.
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