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Cited by 119Muscle Physiology and DisordersMeat and Animal Product QualityViral Infectious Diseases and Gene Expression in Insects

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Development of a Chemically Defined Medium for in vitro Expansion of Primary Bovine Satellite Cells

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Kolkmann, Anna M., Van Essen, Anon, Post, Mark J., Moutsatsou, Panagiota·2022·Frontiers in Bioengineering and Biotechnology
DOI 10.3389/fbioe.2022.895289PMC PMC9385969
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Abstract

The use of fetal bovine serum (FBS) in animal cell culture media is widely spread since it provides a broad spectrum of molecules that are known to support cell attachment and growth. However, the harvest and collection procedures of FBS raise ethical concerns and serum is an ill-defined and expensive component. This is especially problematic when it comes to regulatory approval for food applications like cultured meat. The aim of this study is to develop a chemically defined, cost efficient serum-free and animal-free medium that supports the attachment and expansion of bovine myoblasts while maintaining their differentiation capacity. Bovine satellite cells were harvested and isolated from a fresh sample of skeletal muscle tissue and cultured in planar systems. The efficacy of the tested formulations was assessed with metabolic assays and cell counting techniques. Optical microscopy was used to observe cellular morphology and statistical analysis was applied. Based on a comprehensive literature analysis, a defined serum-free medium (SFM) composition was developed consisting of DMEM/F12 as basal medium, supplemented with L-ascorbic acid 2-phosphate, fibronectin, hydrocortisone, GlutaMAX™, albumin, ITS-X, hIL-6, α-linolenic acid, and growth factors such as FGF-2, VEGF, IGF-1, HGF, and PDGF-BB. To our knowledge, this is the first defined serum-free and animal free medium formulation specific for bovine myoblasts to date. We conclude that the SFM formulation supported exponential cell growth up to 97% of the serum—containing golden standard growth medium. All reagents used in this study are chemically defined.

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References 60 · Cited by 119

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Metadata

Authors
Kolkmann, A. M., Van Essen, A., Post, M. J., & Moutsatsou, P.
Affiliations
Maastricht University (NL)
Corresponding author
Moutsatsou, Panagiota
ORCID
Venue
Frontiers in Bioengineering and Biotechnology
Publisher
Frontiers Media SA
Published
2022-08-04
Volume
10
Pages
895289-895289
ISSN
2296-4185 (ISSN-L)
DOI
10.3389/fbioe.2022.895289
PMC
PMC9385969
OpenAlex
W4289731152
Cited by
119
References
60
Citations by year
2026: 27 · 2025: 34 · 2024: 33 · 2023: 20 · 2022: 5
FWCI
7.51
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Bioengineering and Biotechnology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research Publications (Maastricht University) (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Research Publications (Maastricht University) (submittedVersion, other-oa)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
  • Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Meat and Animal Product Quality
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
Keywords
chemically defined medium, serum-free medium, bovine satellite cells, cultured meat, skeletal muscle, cell attachment, myogenesis, growth factor supplementation, fetal bovine serum alternatives

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