| Autor: |
N. Jenkins, Victor Vinci, G. Sullivan, R. J. Strobel, M. Bracey, D. Bowden, C. Hatfield |
| Rok vydání: |
2001 |
| Předmět: |
|
| Zdroj: |
Animal Cell Technology: From Target to Market ISBN: 9789401038973 |
| DOI: |
10.1007/978-94-010-0369-8_70 |
| Popis: |
Cultivation of animal cells for development of new therapeutic proteins has roudnely employed shaken flasks or roller bottles. In this study, cell cultures were successfully miniaturized in a microtiter plate format and protocols were devised to grow cells in universal 96 deep-well microplates with shaking. Cultivation of CHO DG44 cells expressing recombinant human protein 1 (rhPl) was shown to be equivalent to growth in shaken flasks with respect to rhPl titer and productivity, integral cell area, and metabolite profiles (e.g., lactate and ammonia). Adjustments to the culture volume in microplate wells were found necessary to provide adequate mixing of cell suspensions. Microplate work was expanded to include transformed human kidney cells (HEK293H) as well as AVI2 cells derived from Syrian hamster. Response surface methods were used to assess the similarity of results between vessel types. Shaken microplates served as acceptable substitutes for shaken flasks in all three of the cell lines based on response surface mapping of integral cell area or product titer. The potential of microplates as growth vessels for high throughput process development, media development, and clonal isolation/evaluation is remarkable ∔ 10,000 microplate wells can be used to grow cells in the same amount of laboratory incubator space that would normally beoccupied by only 160 flasks or roller bottles. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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