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Catalogue No.


Cell Line Name


Cell Line Description

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).



None specified


None specified

Tissue of Origin



Not specified


None Stated

Culture Conditions

Subculture Routine

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.

Culture Medium

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

Growth Mode


Additional Info


Dr. R. Dowling, CAMR, Salisbury

Country of Origin

United Kingdom

Hazard Group (ACDP)

Hazard Group (ACDP) 2



PUCK TT, CIECIURA SJ, ROBINSON A 1958 Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects.J Exp Med. 108(6):945-56 PMID: 13598821.

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: CHO (PROTEIN FREE) (ECACC 00102307).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.