Iranian Biomedical Journal

en Optimization of Culture Conditions to Improve Follicle-Stimulating Hormone Production by CHO-DG44 Cells in Serum-Free Medium Pharmaceutical Biotechnology Pharmaceutical Biotechnology مقاله کامل Full Length/Original Article <div style="text-align: justify;">Background: In the present study, we attempted to adapt an adherent and serum-dependent Chinese hamster ovary DG44 cell line to a serum-free suspension culture and optimize the culture condition to achieve a higher yield of recombinant human follicle stimulating hormone (r-hFSH) with acceptable quality. This approach helps to mitigate the risks associated with blood-borne pathogens, reduces lot-to-lot variability, and lowers costs, making it suitable for industrial processing and scale-up. Methods: The cell adaptation was performed using different chemically defined SFM. This process was followed by optimization through statistical experimental design, focusing on selected physicochemical parameters, including chemical supplementation of the medium and temperature shift. Both small- and large-scale cultures were conducted to test the reproducibility of the optimized condition. The expressed protein was evaluated for comparability with the standard molecule according to the Pharmacopeia guidelines. Results: response surface methodology (RSM) analysis indicated that supplementation of the culture medium with galactose and sodium butyrate (NaBu), along with a temperature downshift, were the main parameters leading to increased cell viability (10%), r-hFSH level (96%), and more importantly, the glycosylation content (49%) of r-hFSH compared to the control condition. Conclusion: As r-hFSH isoforms generated during in vivo post-translational modifications typically exhibit different serum/plasma half-lives and bioactivity due to their incorporated sialic acid content/glycosylation, further optimizations of r-hFSH production are necessary to enhance its biological activity. In this study, following a primary screening of the studied parameters, optimization of culture conditions based on selected parameters resulted in enhanced quality and quantity of the produced r-hFSH. However, further examination is necessary before transitioning to industrial production.  </div> CHO cells, Human follicle stimulating hormone, Serum-free culture medium 282 296 http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1933-1&slc_lang=en&sid=1 Hanna Ghobadian hanna.ghobadian@gmail.com 0009-0003-0857-8624 No Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran Khashayar Roshanzamir Roshanzamirk@cinnagen.com 0009-0005-9426-5433 No CinnaGen Medical Biotechnology Research Center, Alborz University of Medical Sciences, Karaj, Iran Mohammadhasan Kouhi Abdolabadi kouhim@cinnagen.com 0009-0003-0592-2450 No CinnaGen Medical Biotechnology Research Center, Alborz University of Medical Sciences, Karaj, Iran Hadi Ostadi ostadih@cinnagen.com 0009-0005-3116-9073 No CinnaGen Medical Biotechnology Research Center, Alborz University of Medical Sciences, Karaj, Iran Reza Zati Keikha zatir@cinnagen.com 0009-0003-7939-8946 No CinnaGen Medical Biotechnology Research Center, Alborz University of Medical Sciences, Karaj, Iran Mohammad Dolatkhah Baghan dolatkhahm@cinnagen.com 0009-0002-6652-5814 No CinnaGen Medical Biotechnology Research Center, Alborz University of Medical Sciences, Karaj, Iran Yeganeh Talebkhan ytgaroosi@gmail.com 0000-0003-1988-4419 Yes Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran Fatemeh Torkashvand fatemehtorkashvand4@gmail.com 0009-0000-1702-4509 No Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran