Merck, 08062569-DNA-5UG, 108CC5-BU-8

Application
Used in the generation of cell hybrids.
Biochem/physiol Actions
A bromodeoxyuridine resistant mutant of 108CC5 (Sigma Catalogue number 08062521), a mouse neuroblastoma N18TG2 x rat glioma C6-BU-1 hybrid
Mouse neuroblastoma x Rat glioma hybrid
Preparation Note
DMEM + 2 mM glutamine + 10% Foetal Bovine Serum (FBS) + 1 mM 8-bromodeoxyuridine. The cells can be grown in the absence of bromodeoxyuridine.
Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 1-2x10,000 cells/cm² using 0.05% trypsin/EDTA; 5% CO₂; 37°C. The cultures rapidly produce lactic acid through anaerobic glycolysis causing the media to become acidified. The medium must never be allowed to become acidic as cells detach in clumps and take many passages to recover. Frequent inspection of cultures is therefore required. Immediately after resuscitation a pre-centrifugation step to remove cryoprotectant (DMSO) is recommended i.e. pellet the cells by centrifugation at 100 x g for 5 minutes and resuspend the cell pellet in fresh medium using the appropriate volume to achieve the correct seeding density.
Other Notes
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This cell line is part of the European Collection of Authenticated Cell Cultures (ECACC), an international repository managed by the United Kingdom Health Security Agency (UKHSA). No licensing agreement is required when either this cell line or the DNA extracted from it are used for internal research purposes only. Any other use of these products is prohibited without the express written permission of UKHSA. Inquiries regarding authorized use of this cell line or its genomic DNA may be directed to [email protected].