Satellite cells sourced from bull calves and dairy cows differs in proliferative and myogenic capacity – Implications for cultivated meat

TY - JOUR

T1 - Satellite cells sourced from bull calves and dairy cows differs in proliferative and myogenic capacity – Implications for cultivated meat

AU - Skrivergaard, Stig

AU - Krøyer Rasmussen, Martin

AU - Sahebekhtiari, Navid

AU - Feveile Young, Jette

AU - Therkildsen, Margrethe

N1 - Publisher Copyright: © 2023 The Authors

PY - 2023/11

Y1 - 2023/11

N2 - Cultivated meat produced with primary muscle satellite cells (SCs) will need a continuous supply of isolated cell material from relevant animal donors. Factors such as age, sex, and breed, along with the sustainability and availability of donor animals, could determine the most appropriate donor type for an efficient production. In this study, we focus on the proliferation and differentiation of bovine SCs isolated from bull calf and dairy cow muscle samples. The proliferative performance of bull calf SCs was significantly better than SCs from dairy cows, however a dynamic differentiation assay revealed that the degree of fusion and formation of myotubes were similar between donor types. Furthermore, the proliferation of SCs from both donor types was enhanced using an in-house developed serum-free media compared to 10% FBS, which also delayed myogenic differentiation and increased final cell population density. Using gene chip transcriptomics, we identified several differentially expressed genes between the two donor types, which could help explain the observed cellular differences. This data also revealed a high biological variance between the three replicate animals within donor type, which seemed to be decreased when using our in-house serum-free media. With the use of the powerful imaging modalities of Cytation 5, we developed a novel high contrast brightfield-enabled label-free myotube quantification method along with a more efficient end-point fusion analysis using Phalloidin-staining. The results give new insights into the bovine SC biology and potential use of bull calves and dairy cows as relevant donor animals for cultivated beef cell sourcing. The newly developed differentiation assays will further enhance future research within the field of cultivated meat and SC biology.

AB - Cultivated meat produced with primary muscle satellite cells (SCs) will need a continuous supply of isolated cell material from relevant animal donors. Factors such as age, sex, and breed, along with the sustainability and availability of donor animals, could determine the most appropriate donor type for an efficient production. In this study, we focus on the proliferation and differentiation of bovine SCs isolated from bull calf and dairy cow muscle samples. The proliferative performance of bull calf SCs was significantly better than SCs from dairy cows, however a dynamic differentiation assay revealed that the degree of fusion and formation of myotubes were similar between donor types. Furthermore, the proliferation of SCs from both donor types was enhanced using an in-house developed serum-free media compared to 10% FBS, which also delayed myogenic differentiation and increased final cell population density. Using gene chip transcriptomics, we identified several differentially expressed genes between the two donor types, which could help explain the observed cellular differences. This data also revealed a high biological variance between the three replicate animals within donor type, which seemed to be decreased when using our in-house serum-free media. With the use of the powerful imaging modalities of Cytation 5, we developed a novel high contrast brightfield-enabled label-free myotube quantification method along with a more efficient end-point fusion analysis using Phalloidin-staining. The results give new insights into the bovine SC biology and potential use of bull calves and dairy cows as relevant donor animals for cultivated beef cell sourcing. The newly developed differentiation assays will further enhance future research within the field of cultivated meat and SC biology.

KW - Bovine satellite cell sourcing

KW - Cell proliferation

KW - Cellular agriculture

KW - Donor type

KW - Image-based cytometry

KW - Label-free quantification

KW - Myogenic differentiation

KW - Transcriptomics.

UR - https://www.scopus.com/pages/publications/85164996027

U2 - 10.1016/j.foodres.2023.113217

DO - 10.1016/j.foodres.2023.113217

M3 - Journal article

C2 - 37803537

AN - SCOPUS:85164996027

SN - 0963-9969

VL - 173

JO - Food Research International

JF - Food Research International

IS - Part 1

M1 - 113217

ER -