Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture

ABSTRACT: In vitro meat cultivation, a cutting-edge innovation in food science, may represent a more sustainable and ethical source of animal proteins compared with conventionally grown meat. An important challenge for meat cultivation lies in eliminating the use of fetal bovine serum (FBS) in cell...

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Main Authors: T.S. Sundaram, C. Giromini, R. Rebucci, D. Lanzoni, E. Petrosillo, A. Baldi, F. Cheli
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Journal of Dairy Science
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Online Access:http://www.sciencedirect.com/science/article/pii/S0022030225000888
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author T.S. Sundaram
C. Giromini
R. Rebucci
D. Lanzoni
E. Petrosillo
A. Baldi
F. Cheli
author_facet T.S. Sundaram
C. Giromini
R. Rebucci
D. Lanzoni
E. Petrosillo
A. Baldi
F. Cheli
author_sort T.S. Sundaram
collection DOAJ
description ABSTRACT: In vitro meat cultivation, a cutting-edge innovation in food science, may represent a more sustainable and ethical source of animal proteins compared with conventionally grown meat. An important challenge for meat cultivation lies in eliminating the use of fetal bovine serum (FBS) in cell culture media due to ethical concerns. Milk whey is a nutrient-rich liquid portion of the milk, derived as a byproduct of dairy industry. Similar to FBS, whey contains proteins that are crucial for nutrition, cell adhesion, and biomolecular transport. In this study, we investigated whether whey proteins (WP) can replace FBS in supporting muscle cell cultivation, using the C2C12 myoblast model. Accordingly, under serum-free conditions, cells were treated with 2 WP mixtures, grouping high (β-LG 1.25%, α-LA 1.25%, BSA 1.25%) and low (β-LG 0.07%, α-LA 0.15%, BSA 0.15%) selected concentrations of individual proteins that positively affected cell growth in a preceding dose–response study. Cells cultured in only basal Dulbecco's Modified Eagle Medium were included as a negative control, and cells cultured in 10% FBS as a positive control. Cells were maintained in the treatment media for 48 h (d 1 and 2) to support myoblast proliferation. Subsequently, all the treatments were replaced with a standard low mitogenic 2% horse serum (HS) medium until full differentiation (d 6). The treatment effects on morphology, viability, and lactate dehydrogenase release were assessed after d 1, 2, and 6, respectively. The results showed that WP stimulated cell proliferation under serum-free culture conditions, similar to the FBS control, and subsequently facilitated myotube formation when the WP or FBS treatments were switched to HS medium. After differentiation, these cells also exhibited increased expression of cell differentiation markers such as creatine kinase and citrate synthase and underwent morphological changes from spindle-shaped cells to fused elongated myotubes, in contrast to the negative control. This study demonstrates that WP are a promising and sustainable alternative for considerably replacing FBS-based growth supplements for use in cultivated animal products.
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spelling doaj-art-0233f25b4e4847dd823805641f9734c52025-08-20T03:13:49ZengElsevierJournal of Dairy Science0022-03022025-05-0110854749476010.3168/jds.2024-25449Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell cultureT.S. Sundaram0C. Giromini1R. Rebucci2D. Lanzoni3E. Petrosillo4A. Baldi5F. Cheli6Department of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, Italy; Corresponding authorDepartment of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, Italy; Institute for Food, Nutrition and Health, University of Reading, Reading RG6 5EU, United Kingdom; Innovation for Well-Being and Environment (CRC I-WE), University of Milan, 20122 Milan, ItalyDepartment of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, ItalyDepartment of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, ItalyDepartment of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, ItalyDepartment of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, ItalyDepartment of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, Italy; Innovation for Well-Being and Environment (CRC I-WE), University of Milan, 20122 Milan, ItalyABSTRACT: In vitro meat cultivation, a cutting-edge innovation in food science, may represent a more sustainable and ethical source of animal proteins compared with conventionally grown meat. An important challenge for meat cultivation lies in eliminating the use of fetal bovine serum (FBS) in cell culture media due to ethical concerns. Milk whey is a nutrient-rich liquid portion of the milk, derived as a byproduct of dairy industry. Similar to FBS, whey contains proteins that are crucial for nutrition, cell adhesion, and biomolecular transport. In this study, we investigated whether whey proteins (WP) can replace FBS in supporting muscle cell cultivation, using the C2C12 myoblast model. Accordingly, under serum-free conditions, cells were treated with 2 WP mixtures, grouping high (β-LG 1.25%, α-LA 1.25%, BSA 1.25%) and low (β-LG 0.07%, α-LA 0.15%, BSA 0.15%) selected concentrations of individual proteins that positively affected cell growth in a preceding dose–response study. Cells cultured in only basal Dulbecco's Modified Eagle Medium were included as a negative control, and cells cultured in 10% FBS as a positive control. Cells were maintained in the treatment media for 48 h (d 1 and 2) to support myoblast proliferation. Subsequently, all the treatments were replaced with a standard low mitogenic 2% horse serum (HS) medium until full differentiation (d 6). The treatment effects on morphology, viability, and lactate dehydrogenase release were assessed after d 1, 2, and 6, respectively. The results showed that WP stimulated cell proliferation under serum-free culture conditions, similar to the FBS control, and subsequently facilitated myotube formation when the WP or FBS treatments were switched to HS medium. After differentiation, these cells also exhibited increased expression of cell differentiation markers such as creatine kinase and citrate synthase and underwent morphological changes from spindle-shaped cells to fused elongated myotubes, in contrast to the negative control. This study demonstrates that WP are a promising and sustainable alternative for considerably replacing FBS-based growth supplements for use in cultivated animal products.http://www.sciencedirect.com/science/article/pii/S0022030225000888whey proteinsfetal bovine serum alternativecultured meatcellular food
spellingShingle T.S. Sundaram
C. Giromini
R. Rebucci
D. Lanzoni
E. Petrosillo
A. Baldi
F. Cheli
Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
Journal of Dairy Science
whey proteins
fetal bovine serum alternative
cultured meat
cellular food
title Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
title_full Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
title_fullStr Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
title_full_unstemmed Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
title_short Milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
title_sort milk whey as a sustainable alternative growth supplement to fetal bovine serum in muscle cell culture
topic whey proteins
fetal bovine serum alternative
cultured meat
cellular food
url http://www.sciencedirect.com/science/article/pii/S0022030225000888
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