In vitro culture of bovine fibroblasts using select serum-free media supplemented withChlorella vulgarisextract

Abstract

Standard cell culture practices require addition of animal-derived serum to culture media to achieve adequate cell growth. Typically, 5-10% by volume of fetal bovine serum (FBS) is used, which accounts for a vast majority of the cost of media while also imposing environmental and ethical concerns associated with the use of animal serum. Here we tested the efficacy of culturing cells by replacing serum in the media with algae extract and select additives. Using LC-MS, we compared molecular signatures of FBS to Chlorella algae extracts and identified NAD(H)/NADP(H) as common and relatively abundant features in their characteristic profiles. Bovine fibroblasts, cultured in serum-free media supplemented with C. vulgaris extract and just two growth factors plus insulin, showed significant growth with enhanced viability compared to control cells cultured without serum, albeit still lower than that of controls cultured with 10% FBS. Moreover, C. vulgaris extract enhanced cell viability beyond that of cells cultured with the two growth factors and insulin alone. These results suggest that key components in serum which are essential for cell growth may also be present in C. vulgaris extract, demonstrating that it may be used at least as a partial alternative to serum for cell culture applications.

Evaluating the Potential of Marine Invertebrate and Insect Protein Hydrolysates to Reduce Fetal Bovine Serum in Cell Culture Media for Cultivated Fish Production

“…Lower concentrations of whey protein hydrolysate (0.01–0.5 mg/mL) boosted osteoblastic cell line proliferation, viability, and alkaline phosphatase activity [ 63 ]. Similar results were observed on bovine cells with algal extract and various other industrial byproduct hydrolysates; the cell growth increased relative to the serum-free conditions, albeit less than in the serum-rich conditions [ 13 , 64 ].…”

Evaluating the potential of marine invertebrate and insect protein hydrolysates to reduce fetal bovine serum in cell culture media for cultivated fish production

“…Lower concentrations of whey protein hydrolysate (0.01-0.5 mg/mL) boosted osteoblastic cell line proliferation, viability, and alkaline phosphatase activity (Jo et al, 2020). Similar results were observed on bovine cells with algal extract and various other industrial byproduct hydrolysates; cell growth increased relative to serum-free conditions, albeit less than in serum-rich conditions (Andreassen et al, 2020; Defendi-Cho & Gould, 2021).…”