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Short- and long-term effects on mAb-producing CHO cell lines after cryopreservation
Jayashree Subramanian1, Rigzen P S Aulakh1, Parbir S Grewal1
1Early Stage Cell Culture, Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, CA, 94080.
Biotechnology Progress
|January 10, 2018
Summary
Cryopreservation does not harm Chinese hamster ovary (CHO) cell lines, even after thawing. However, cell-specific productivity (Qp) declines with cell age, highlighting the need for clone selection and mechanism studies.
Area of Science:
- Biotechnology
- Cell Biology
- Biopharmaceutical Manufacturing
Background:
- Cryopreservation is crucial for Chinese hamster ovary (CHO) cell biopharmaceutical manufacturing.
- Limited data exist on cryopreservation's impact on CHO cell lines and their productivity over time.
- The glutamine synthetase (GS)/methionine sulfoximine (MSX) selection system is widely used for CHO cell line development.
Purpose of the Study:
- To assess the impact of cryopreservation and thawing on CHO cell line performance.
- To investigate the decline in cell-specific productivity (Qp) over time in recombinant CHO cells.
- To differentiate the effects of cryopreservation from those of cell aging on CHO cell productivity.
Main Methods:
- Evaluated 24 recombinant CHO cell lines using production culture assays.
- Compared performance (growth, viability, productivity) of cryopreserved versus non-cryopreserved cells.
- Analyzed product quality (size, charge, glycosylation) and flow cytometry profiles.
Main Results:
- Freeze-thaw cycles did not significantly impact CHO cell performance beyond initial passages.
- Cryopreserved and non-cryopreserved cells showed similar growth, viability, productivity, and product quality.
- 17 out of 24 cell lines exhibited a ≥20% decline in Qp after 2-3 months, independent of cryopreservation.
Conclusions:
- Cryopreservation is a safe and effective method for CHO cell banking without lasting detrimental effects.
- Cell age, not cryopreservation, is a significant factor in the decline of cell-specific productivity (Qp).
- Further research into Qp decline mechanisms and robust clone selection are essential for biopharmaceutical manufacturing.
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