Cell culture and gene transcription effects of copper sulfate on Chinese hamster ovary cells
| Autor: | Sarwat F. Khattak, Zheng Jian Li, Zizhuo Xing, Paul S. Kayne, Nan-xin Qian, Yueming Qian, Aiqing He, Shih-Hsie Pan |
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| Rok vydání: | 2011 |
| Předmět: |
Copper Sulfate
Recombinant Fusion Proteins Cell Blotting Western Cell Culture Techniques Transferrin receptor CHO Cells Biology chemistry.chemical_compound Lactate dehydrogenase Cricetinae medicine Animals Lactic Acid Microarray analysis techniques Cell growth Chinese hamster ovary cell Cell Cycle Cell cycle Molecular biology medicine.anatomical_structure Biochemistry chemistry Cell culture Immunoglobulin G Biotechnology |
| Zdroj: | Biotechnology progress. 27(4) |
| ISSN: | 1520-6033 |
| Popis: | This study reports the effects of varying concentrations of copper sulfate on the metabolic and gene transcriptional profile of a recombinant Chinese hamster ovary (CHO) cell line producing an immunoglobulin G (IgG)-fusion protein (B0). Addition of 50 μM copper sulfate significantly decreased lactate accumulation in the cultures while increasing viable cell density and protein titer. These changes could be seen from day 6 and became increasingly evident with culture duration. Reducing the copper sulfate concentration to 5 μM retained all the above beneficial effects, but with the added benefit of reduced levels of the aggregated form of the B0 protein. To profile the cellular changes due to copper sulfate addition at the transcriptional level, Affymetrix® CHO microarrays were used to identify differentially expressed genes related to reduced cellular stresses and facilitated cell cycling. Based on the microarray results, down-regulation of the transferrin receptor and lactate dehydrogenase, and up-regulation of a cytochrome P450 family-2 polypeptide were then confirmed by Western blotting. These results showed that copper played a critical role in cell metabolism and productivity on recombinant CHO cells and highlighted the usefulness of microarray data for better understanding biological responses on medium modification. |
| Databáze: | OpenAIRE |
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