RG2

The RG2 cell line is derived from a chemically induced glioma in Fischer 344 rats. Generated via transplacental administration of N-ethyl-N-nitrosourea (ENU), RG2 gliomas are classified as anaplastic gliomas due to their invasive growth pattern, high mitotic index, and undifferentiated morphology. These tumors are notable for their consistent lethality in vivo and their ability to grow in syngeneic hosts without eliciting a significant immune response. This low immunogenicity makes RG2 an ideal model for studying glioblastoma-like tumors and testing experimental therapies in immunocompetent settings.

RG2 glioma cells exhibit characteristics typical of high-grade gliomas, including rapid proliferation, invasive capacity, and genomic alterations. Studies have highlighted the loss of tumor suppressor genes such as CDKN2A, along with dysregulated pathways involving PDGF, Ras, and IGF signaling. The cell line grows as undifferentiated spindle-shaped cells in vitro, maintaining their tumorigenic potential when implanted intracranially, where they display diffuse invasion into normal brain tissue, mimicking human glioblastoma behavior.

This cell line has been extensively utilized in preclinical research to evaluate the efficacy of various therapeutic approaches, including chemotherapy, radiotherapy, gene therapy, and immunotherapy. RG2 gliomas are particularly valuable for testing novel drug delivery methods, such as convection-enhanced delivery (CED), and for investigating mechanisms of blood-brain barrier disruption in gliomas. Its histopathological and molecular resemblance to human glioblastomas underscores its utility in translational neuro-oncology.

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.