Expansion of bovine skeletal muscle stem cells from spinner flasks to benchtop stirred-tank bioreactors for up to 38 days

Dimitrios Tzimorotas; Nina Therese Solberg; R Christel Andreassen; Panagiota Moutsatsou; Vincent Bodiou; Mona Elisabeth Pedersen; Sissel Beate Rønning

doi:10.3389/fnut.2023.1192365

Expansion of bovine skeletal muscle stem cells from spinner flasks to benchtop stirred-tank bioreactors for up to 38 days

Front Nutr. 2023 Aug 7:10:1192365. doi: 10.3389/fnut.2023.1192365. eCollection 2023.

Authors

Dimitrios Tzimorotas¹, Nina Therese Solberg², R Christel Andreassen², Panagiota Moutsatsou³, Vincent Bodiou³, Mona Elisabeth Pedersen², Sissel Beate Rønning²

Affiliations

¹ Nofima AS, Food Safety and Health, Ås, Norway.
² Nofima AS, Raw Materials and Process Optimization, Ås, Norway.
³ Mosa Meat BV, Maastricht, Netherlands.

Abstract

Introduction: Successful long-term expansion of skeletal muscle satellite cells (MuSCs) on a large scale is fundamental for cultivating animal cells for protein production. Prerequisites for efficient cell expansion include maintaining essential native cell activities such as cell adhesion, migration, proliferation, and differentiation while ensuring consistent reproducibility.

Method: This study investigated the growth of bovine MuSC culture using low-volume spinner flasks and a benchtop stirred-tank bioreactor (STR).

Results and discussion: Our results showed for the first time the expansion of primary MuSCs for 38 days in a bench-top STR run with low initial seeding density and FBS reduction, supported by increased expression of the satellite cell marker PAX7 and reduced expression of differentiation-inducing genes like MYOG, even without adding p38-MAPK inhibitors. Moreover, the cells retained their ability to proliferate, migrate, and differentiate after enzymatic dissociation from the microcarriers. We also showed reproducible results in a separate biological benchtop STR run.

Keywords: bioreactor; microcarriers; skeletal muscle satellite cell; stem cell expansion; upscaling.

Grants and funding

We thank the Norwegian Fund for Research Fees for Agricultural Products (FFL) for supporting the study through the project “Precision” (project number 314111).