Investigating the Feasibility of Soy Protein Hydrolysate in Substituting Fetal Bovine Serum in Cancer Cell Culture

Tan, Adrian Zhen Ling (2023) Investigating the Feasibility of Soy Protein Hydrolysate in Substituting Fetal Bovine Serum in Cancer Cell Culture. Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Fetal Bovine Serum (FBS) is widely used in cell and tissue culture systems as a supplementation in culture media due to its high content of growth-promoting factors required for cell growth. FBS is a good source of growth factors, adherence and spreading factors, amino acids, vitamins, lipids, hormones and contains a minimum amount of complement proteins. However, FBS is prone to batch-to-batch variation, possible contaminations such as mycoplasma, viruses, and endotoxins which may affect the quality. Due to increasing demand for FBS globally, the price of FBS also increased by over 300% which leads to the necessity of developing alternative products to substitute FBS in the culture system. In this study, the performance of soy protein hydrolysate (SPH) in serum-free culture media growing human liver cancer cell line, HepG2, was compared with FBS. Papain and pepsin enzymes were used to hydrolyse soy protein isolate (SPI) to yield SPH. The cell viability of HepG2 was examined by MTT assay, in which the RPMI medium containing SPH ranging from 1-5% (w/v) was used to culture HepG2 cells for duration of 24 and 48 hours. The maximum concentration tested, 5% pepsin-treated SPH, was able to increase cell viability of HepG2 cells to 184.1% as compared to the control with 10% FBS. Culturing of HepG2 cells in T25 flasks with medium containing 3% and 5% (w/v) SPH, respectively, revealed that no cell survived during the following passage. In conclusion, mediums with 1-5% (w/v) SPH had comparable growth promoting capabilities to that of FBS up to 48 hours, but were unable to sustain the effect for long term. A more complete hydrolysis method of SPI to obtain a clear solution of SPH is needed in future studies to further validate the potential of SPH in substituting FBS in cancer cell culture