OP9

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.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.