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Recently, genetic technique has developed gradually and applied to bioscience technology. So how to direct microorganisms, plants and mammalian cells to produce recombinant proteins has become an important and researchable field. The purpose of this thesis is to study the utilization of transfected mammalian cells to produce recombinant proteins and design.
Granulocyte macrophage colony-stimulating factor (GM-CSF) transfected mammalian 3T3 cells was selected to be our study target.We found that transfected 3T3 had better growth rate and activity at 37℃,but better recombinant proteins productivity at 33℃.The phenomenon of diminish GM-CSF productivity was also noted in sequent generation 3T3 cells line transfected by vector pcDNA3.Further evaluation of vector pTriEx1.1 Neo 3T3 cells line is processed.
Besides the genetic technique,we also pay much attention to decrease the shear force occurred during culturing in bioreactor and improve the adhesive,differentiated and secretive capacity of mammalian cells by immobilization.
We selected polyurethane (PU) and loofa sponge (LS) as the scaffolds. We find that LS was more proper than PU as a scaffold for 3T3 cells to manufacture recombinant proteins. Finally we would cultivate the transfected 3T3 cells in spinner flask and 3D bioreactor respectively and try and error on controlling the spinner rate and the period of changing medium.
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