Tetrahydrofolate Additive for CHO Cell Culture Growth
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Solution Overview
Problem
Conventional chemically defined media for cell culture require long-term adaptation for suspension culture of CHO DG44 host cells and exhibit low cell growth performance, even after adaptation, due to the lack of tetrahydrofolate (THF) supplementation.
Innovation Solution
Incorporating tetrahydrofolate (THF) or its precursors/derivatives into a chemically defined medium to support cell growth, specifically for DHFR gene-deficient CHO cells, enhancing cell density and reducing doubling time, thereby eliminating the need for suspension adaptation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If chemically defined medium is used for cell culture, then medium consistency and traceability are improved, but cell growth performance deteriorates
Solution Approach 1:
The invention changes the chemical composition parameters of the chemically defined medium by adding tetrahydrofolate (THF) at specific concentrations (0.1-100 μM). This parameter modification resolves the contradiction by enhancing cell growth performance while preserving the consistency and traceability characteristics of chemically defined media.
2Adaptability or versatility
If long-term subculture adaptation is performed, then suspension culture capability is improved, but cell growth performance deteriorates
Solution Approach 1:
The invention applies preliminary action by adding tetrahydrofolate to the medium before cell culture begins. This pre-treatment enables cells to achieve suspension culture capability and maintain high growth performance without requiring long-term adaptation through multiple subculture passages, thus resolving the contradiction between adaptability and productivity.
3Productivity
If tetrahydrofolate is added to chemically defined medium, then cell growth performance is improved, but medium complexity increases
Solution Approach 1:
The invention applies local quality by adding a specific substance (tetrahydrofolate) at a targeted location (in the medium formulation) to address the specific nutritional deficiency of DHFR-deficient cells. This localized modification improves cell growth performance while minimizing overall medium complexity, as only one additional component is introduced to the chemically defined formulation.
Data Source
AI summary
The present invention relates to an optimal medium for growing a cell line auxotrophic for tetrahydrofolate (THF) and producing a desired material in the cell with high efficiency. In particular, the present invention provides a method for enhancing cell growth by adding tetrahydrofolate (THF), or a precursor or derivative thereof into a chemical composition cell medium.