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Dissertation
The effect of growth surface properties on the cultivation of anchorage-dependent animals cells and the development of novel surfaces for cell culture
Doctor of Philosophy (Ph.D.), Drexel University
Dec 1994
DOI:
https://doi.org/10.17918/00009783
Abstract
The physiological responses of anchorage dependent animal cells (3T6 mouse embryo fibroblast and A375 human malignant melanoma) to systematic variation of growth surface properties were determined for sulfonated polystyrene (PS-S), styrene-methacrylic acid copolymers (PS-MAA), and styrene-dimethylaminoethyl methacrylate copolymers (PS-DMAEMA). The optimal surface charge densities for attachment, spreading, and proliferation occur over narrow ranges and are not equal. On PS-S, the optimum charge densities for 3T6 are 2.5 charges/nm2 for attachment, 2.0 charges/nm2 for spreading, and 4.0 charges/nm2 for growth, while the maxima for A375 are at 2.0, 3.5, and 2.0 charges/nm2, respectively. On PS-MAA, the maxima for 3T6 are at 40-60 vol% MAA for attachment, 70 vol% MAA for spreading, and 50-60 vol% MAA for growth, while the maxima for A375 are at 20, 80, and >20 vol% MAA, respectively. On PS-DMAEMA, the maxima for 3T6 are at 70 vol% DMAEMA for both attachment and spreading, while the maxima for A375 are at 90-100 vol% DMAEMA for attachment and 60 vol% DMAEMA for spreading. The influence of inoculum density on cell attachment and growth was also studied. The initial cell density (cells/surface area) must be 103-104 cells/cm2 for 3T6 and 3 x 103 - 4 x 104 cells/cm2 for A375 to compare growth rates on different surfaces. To objectively compare cell attachment rates, the rate should be measured as cells/volume-time and the liquid phase cell concentration must be essentially constant. A fixed-surface bioreactor based on a reticulated vitreous carbon foam was used to cultivate 3T6, providing a high surface-to-volume ratio (17 cm2/cm3) and a low-shear (<0.1 dynes/cm2), chemically-inert. and potentially reusable environment. External medium circulation allowed aeration, nutrient monitoring, and medium replacement without disturbing the cells. Optimal flow rates for attachment (120 mL/h) and growth (>400 mL/h) were determined. Growth rates comparable to agitated microcarrier cultures were achieved.
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Details
- Title
- The effect of growth surface properties on the cultivation of anchorage-dependent animals cells and the development of novel surfaces for cell culture
- Creators
- Bradley Lane Kent
- Awarding Institution
- Drexel University
- Degree Awarded
- Doctor of Philosophy (Ph.D.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Number of pages
- xviii, 338 pages
- Resource Type
- Dissertation
- Language
- English
- Academic Unit
- College of Engineering (1970-2026); Drexel University
- Other Identifier
- 991021888854604721