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ORIGINAL RESEARCH article

Front. Bioeng. Biotechnol.
Sec. Bioprocess Engineering
Volume 12 - 2024 | doi: 10.3389/fbioe.2024.1443914

Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum

Provisionally accepted
Vincent Bodiou Vincent Bodiou 1,2Anitha A. Kumar Anitha A. Kumar 1Edoardo Massarelli Edoardo Massarelli 1Tessa van Haaften Tessa van Haaften 1Mark J. Post Mark J. Post 1,2Panagiota Moutsatsou Panagiota Moutsatsou 1*
  • 1 Mosa Meat BV, Maastricht, Netherlands
  • 2 Faculty of Health, Medicine and Life Sciences, Maastricht University, Maastricht, Netherlands, Netherlands

The final, formatted version of the article will be published soon.

    To bring cultivated beef to the market, a scalable system that can support growth of bovine satellite cells (bSCs) in a serum-free and preferably also animal-free medium is of utmost importance. The use of microcarriers (MCs) is, at the moment, one of the most promising technologies for scaling up. MCs offer a large surface to volume ratio, they can be used in scalable stirred tank bioreactors, where the culture conditions can be tightly controlled to meet the cells' requirements (temperature, pH, dissolved oxygen). The inherent capacity of the cells to migrate from one MC to another, also known as bead-to-bead transfer, facilitates a scale-up strategy involving MCs. Previous studies have shown growth of bSCs on three commercially available MCs in serum containing media. Unfortunately there is currently no information available regarding their growth on MCs in serum-free conditions. In this study, we aimed to find suitable serum-free media, MCs and attachment promoting compounds (APCs) supporting the growth of bSCs. Initially, six commercial MCs and three serum-free media were evaluated.The effects of three APCs were compared (vitronectin, laminin and fibronectin). Subsequently, the effects of different concentrations and modes of addition of the best performing APC were investigated. Our results showed that Cytodex 1, Synthemax II and CellBIND supported bSCs' growth in all serum-free media. Overall, better growth was observed with Cytodex 1 in serumfree proliferation media. We showed that the use of laminin or vitronectin with Cytodex 1 can significantly improve cell growth and purity. Laminin also allowed attachment and growth of bSCs on Plastic MCs which had been previously unsuccessful without APCs. Finally, we optimized the use of vitronectin from a sustainability and process perspective, and showed that it can be used solely as a coating for Cytodex 1 (16-100 ng/cm2 ) MCs, instead of as a medium supplement, enhancing cell attachment and proliferation.

    Keywords: cultivated meat, bovine myoblasts, Microbeads, cell expansion, proliferation, bioprocessing, cell attachment, coating

    Received: 04 Jun 2024; Accepted: 16 Oct 2024.

    Copyright: © 2024 Bodiou, Kumar, Massarelli, van Haaften, Post and Moutsatsou. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Panagiota Moutsatsou, Mosa Meat BV, Maastricht, Netherlands

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.