P-124: Comparative Study of The Effect of Fetal Bovine Serum Concentrations on Expressing of Human Follicle Stimulating Hormone.
Follicle stimulating hormone (FSH) produced in recombinant Chinese hamster ovary (CHO) cells used for infertility treatment. Fetal Bovine Serum (FBS) in known as a factor in medium culture that provides the tolerance towards physiochemical variations and protects cells against apoptosis; on the other hand, FBS is the most expensive component in medium culture and collection that cause unnecessary suffering for the unborn calf. This study tried to investigate the effect of FBS levels on growth of recombinant cells producing FSH and FSH titer for determination the optimum concentration of FBS in medium culture.
Materials and methods
Trypan blue staining method was applied for cell counting and measuring Cells Viability. SDS-PAGE and Western blotting used for detection of rFSH. In continuous, qRT-PCR and ImageJ software were used for quantification assays.
The obtained results in this study showed that the most productivity of the cell at 10% FBS. Also, Increase in cell viability and longevity was indicated at 3% FBS. Quantification assays demonstrated reduction in the expression of FSH following the decline in the concentration of FBS. No significant difference was found in the expression of protein at 3 and 5% of FBS.
The maximum expression was seen at medium culture with 10% FBS. Cells viability and longevity increased in medium containing 3% FBS, also the need for FBS reduced to less than one-third in medium with 3% FBS. Considering these, medium containing 3% FBS suggested in this study.