Optimization of Key Factors in Serum Free Medium for Production of Human Recombinant GM-CSF Using Response Surface Methodology

Document Type: Research article


1 Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.


Researchers add serum to a classical medium at concentrations of 5 to 10% (v/v) to grow cells
in-vitro culture media. Unfortunately, serum is a poorly defined culture medium component
as its composition can vary considerably while serum-free cell culture media are an excellent
alternative to standard serum-containing media and offer several major advantages. Advantages
of using serum-free media include a lower risk of infectious agents, lower risk of interfering
components, less contaminant, avoids ethical issues. According to previous studies insulin,
selenium, transferrin and glucose are important component of serum that affect cell growth. In
the present study, we optimized amount of these factors in order to serum free culture medium
fabrication. Response surface methodology (RSM) was employed for optimization of key factors
in serum free medium to enhance recombinant human GM-CSF (rhGM-CSF) production in CHO
cell line. Four important process parameters including insulin concentration (0-2 g/L), transferrin
concentration (0-1 g/L), selenium concentration (0-0.001 g/L) and glucose concentration (0-5
g/L) were optimized to obtain the best response of rhGM-CSF production using the statistical
Box–Behnken design. The experimental data obtained were analyzed by analysis of variance
(ANOVA) and fitted to a second-order polynomial equation using multiple regression analysis.
Numerical optimization applying desirability function was used to identify the optimum
conditions for maximum production of rhGM-CSF. The optimum conditions were found to be
insulin concentration = 1.1 g/L, transferrin concentration = 0.545 g/L, selenium concentration =
0.000724 g/L and glucose = 1. 4 g/L. Maximum rhGM-CSF production was found to be 3.5 g/L.


Main Subjects