CD CHO Medium

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	CD-CHO medium containing 8 mM L-glutamine

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CD-CHO medium containing 8 mM L-glutamine

Publication protocol

A donor parental CHO-S cell line (Life Technologies, Paisley, UK) was used to isolate 22
clonally-derived cell lines using two rounds of limited dilution cloning (LDC) in 96-well plates
as described previously (Davies et al. 2012), yielding a probability of clonality of 0.97 and a
cloning efficiency of 1.15%. After 63 days of expansion in static culture in CD-CHO medium
containing 8 mM L-glutamine – with a sequential scaling up from 96-well plate into a 24-well
plate, 6-well plate, and T-75 cell culture flask, 22 clonally-derived cell lines were transferred
into individual 125 mL Erlenmeyer suspension culture flasks, maintained at 37°C, 140 rpm
under 5% CO2 and cultured until cell viability was recovered and maintained above 95% for
three consecutive passages. At this point, generation number was set at zero and cells were
sampled for cell banking.


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