CHO Protein-Free Cell Line from Chinese hamster ovary cells

A subclone of the parental CHO line originated by Puck in 1957. The cells have an absolute requirement for L-proline. The cells have been adapted for growth in Sigma CHO Protein Free Medium (C5467). They have been cryopreserved in Sigma Cell Freezing Medium - Serum Free (C2639).

Chinese hamster ovary

Sigma CHO-protein free medium + 4mM Glutamine. Cells should be frozen in either Sigma Cell Freezing Medium - Serum Free (C2639) with 10% DMSO or Sigma CHO Protein Free Medium (C5467) with 7.5%-10% DMSO

Viability may be poor on resuscitation and may initially decrease further. Full recovery may take up to 2 weeks. A centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 10⁵ cells/ml. Leave culture flask upright and observe regularly until viable proliferating cells are seen. Once established use a split ratio of 1:2 approximately every 4 to 5 days; 5% CO₂; 37°C.

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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 culturecollections@ukhsa.gov.uk.