Marine-Derived Extracts as Economically Sustainable and Ethical Alternatives to Fetal Bovine Serum in HEK293 Cell Culture

Abstract

Fetal Bovine Serum (FBS) is a staple supplement in cell culture, providing key growth factors that promote cell health and proliferation. Since 1885, cell culture has been a cornerstone of both basic and applied research, leading to significant discoveries of cell communication and function. However, the use of FBS presents economic and ethical challenges. Millions of bovine fetuses are euthanized annually to produce the volume of FBS necessary for in vitro research. Batch-to-batch inconsistencies often hinder experiment replicability, and FBS can account for over 75% of cell culture costs. It is critical to identify viable FBS alternatives for the sustainability of in vitro research. Marine sources offer an avenue as FBS alternatives because of their abundance of bioactive compounds and sustainable potential. In this study, novel extracts derived from sea urchin roe (known as uni) and the algal species Gracilaria salicornia invasive to Hawaiʻi, were evaluated as alternatives to FBS. Extracts were prepared at 5%, 10%, and 20% concentrations in Dulbecco’s Modified Eagle Medium (DMEM) and exposed to Human Embryonic Kidney (HEK293) cells in culture for 3 days. Cell proliferation, viability, and morphology were assessed through cell quantification, MTT assays, and microscopy. Cells grown in 5% and 10% uni extract and 10% G. salicornia extract treatments showed promising proliferation, with cell counts statistically comparable (p>0.05) to the FBS control. Cells grown in the 5% uni treatment exhibited the healthiest morphology, comparable to 84.3% of the cell population in the FBS control group. Despite the proliferative success, the MTT assay showed significantly lower viability across all treatments (p<0.001), suggesting the extracts may lack metabolic factors necessary for cellular function. These findings highlight the limited potential of marine-derived extracts as FBS supplements and provide a platform for continued exploration and improvement in the field.