| Summary: | 碩士 === 大同大學 === 生物工程學系(所) === 96 === Glycosylation and post-translational modification of recombinant proteins are the unique feature of mammalian cell system. However, the productivity of recombinant protein by mammalian cells is much lower than that of microorganisms. Chinese hamster ovary cell line is the most popular mammalian host for the commercial production of therapeutic proteins. We used Chinese hamster ovary cells (5/9 M alpha 3-18, BCRC 60185, CHO cells) to express macrophage colony-stimulating factor (M-CSF), which can stimulate macrophage proliferaction, differenciation and survival.
Cells were inoculated into culture system containing light weight porous microspheres, which were made of biodegradable polymer, PLGA. The support of PLGA microspheres reduced cell damage resulting from the shear force.
The particle size of dense PLGA particles was 117.8±15.0 μm；1% NH4HCO3-PLGA microcarrier was about 375.8±78.5 μm；5 % NH4HCO3-PLGA particle size was 442.4±25.8μm。The pore size of 1 % NH4HCO3-PLGA and 5 % NH4HCO3-PLGA was between 17~18μm。
Cytodex 3 is a commercial cell culture microcarrier, which was cross-linked dextran coated with denatured collagen. After cell culture with Cytodex 3 for 4 days, the cell density reached maximum, and the onset of cell death was observed. The cell culture with PLGA microcarriers, made of 1% NH4HCO3 and PLGA, the cells grew till day 13. The final density of cells was similar to that cultured with Cytodex 3. The 5 % NH4HCO3-PLGA microcarriers showed the massive cell death up to day 17, but the cell growth rate and cell density were both smaller to that with 1% PLGA microcarriers. The yield of M-CSF is higher for cell culture with 1 % NH4HCO3-PLGA microcarrier. The 1% NH4HCO3-PLGA microcarrier was the best in cell density and M-CSF production.
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