# Effect of whey protein hydrolysate isolation as a partial replacement of fetal bovine serum on proliferation and differentiation of Hanwoo primary muscle cells

## TL;DR

This study shows that whey protein hydrolysate can replace fetal bovine serum in growing Hanwoo muscle cells, reducing costs and ethical concerns.

## Contribution

The study introduces whey protein hydrolysate as a sustainable alternative to fetal bovine serum in muscle cell culture.

## Key findings

- WPH at 0.9 mg/mL supported cell proliferation similar to 20% FBS.
- WPH at 0.06 mg/mL enhanced differentiation with higher MyHC expression than FBS.
- WPH offers a cost-effective and ethical replacement for FBS in cultured meat production.

## Abstract

This study aimed to evaluate the potential of whey protein hydrolysate isolation (WPH) to partially replace fetal bovine serum (FBS) in Hanwoo primary muscle cell cultures, and to address issues related to cost, animal ethics, and undefined elements of FBS. WPH was prepared by enzymatic hydrolysis using bromelain. For proliferation, 10% FBS and 0.3–0.9 mg/mL WPH were used, and for differentiation, 1% FBS and 0.03–0.09 mg/mL WPH were applied. The results showed that cell proliferation increased with increasing WPH concentration, and the levels were similar to those of 20% FBS on days 4 and 5. Therefore, 0.9 mg/mL WPH was considered the most suitable concentration for proliferation. Differentiation initially increased and then decreased with increasing WPH concentration, but mRNA MyHC expression was higher than that of the 20% FBS group. Therefore, 0.06 mg/mL WPH was found to be the most suitable concentration for differentiation. These results suggest that WPH can effectively replace FBS to reduce the production costs of cultured meat and increase the value of by-product utilization, while providing a sustainable and ethical alternative source of nutrients.