Modulation of HEK293T cells proliferation under different conditions by c-Myc overexpression.
Mammalian cells are widely used to produce biopharmaceutical products. However, mammalian cells are slow in development, sensitive to nutrient lost and metabolic wastes accumulation. Hence, proliferation and viability becomes the main features in achieving high protein yield. This can be achieved th...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2013
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| Online Access: | http://hdl.handle.net/10356/52913 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | Mammalian cells are widely used to produce biopharmaceutical products. However, mammalian cells are slow in development, sensitive to nutrient lost and metabolic wastes accumulation. Hence, proliferation and viability becomes the main features in achieving high protein yield. This can be achieved through genetic engineering in which c-myc is overexpressed in HEK293T cells. Overexpression of c-Myc leads to higher cell number complete DMEM. Under low glucose and serum conditions, c-Myc overexpression resulted in higher cell number and viability than untransfected cells. However, when cells were glucose starved, cell viability decreased. Cell number and viability of c-Myc transfected cells were lower than untransfected cells which indicate a stronger dependency on glucose. When cells were serum starved, c-Myc overexpression reduced cell death rate. Effects of c-Myc overexpression were shown to be strongest in high lactate and ammonium DMEM. c-Myc transfected cells had cell number that is 2 fold higher in high lactate concentration and 3 fold higher in high ammonium concentration when compared to untransfected cells. In conclusion, overexpressing c-Myc shows potential to generate a robust cell line that has increase proliferation and reduced cell death. |
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