Past Issue

Volume 9, Supplement 1, Summer 2015 (Presented at 16th Congress on Reproductive Biomedicine and 10th Royan Nursing and Midwifery Seminar) Pages: 84-84

P-98: The Effect of Primary pH Values of Medium Culture on Expression of Human Follicle Stimulating Hormone in Recombinant Chinese Hamster Ovary Cells.


Background
One of the common gonadotropin hormones used for infertility treatment is recombinant Follicle stimulating hormone (rFSH). pH is an environmental factor that can affect production of recombinant proteins with no imposing cost. Culture pH is known as a parameter that significantly affects cell growth and protein production. In this study, tried to investigate the effect of different pH values on cells growth and rFSH production.��
Materials and methods
The primary pH changes were carried out by adding NaOH and HCL 1 N to medium culture. The range of considered pH values was (6.7-7.6). Trypan blue staining method was applied for cells counting and measuring viability of the cells. Total protein concentration was determined by Bradford assay and Western blotting used for detection of rFSH. qRT-PCR and ImageJ software were used for quantification assays.
Results
According to obtained data, at the first stationary phase of cells growth, by evaluating the pH values to 7/6, the cells viability, mRNA ratio of rFSH and rFSH production significantly was decreased. The highest cells viability and mRNA ratio of FSH and FSH production were seen in medium with pH=7.0. The achieved results from relative gene expression at pH=6.7 and pH=7.0 didn�t show a significant difference.
Conclusion
Considering obtained data, by lowering pH value to 7.0 for cells, without any cost, the highest proliferation, will be achieved.