Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum
IntroductionTo 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...
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Frontiers Media S.A.
2024-11-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1443914/full |
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| author | Vincent Bodiou Vincent Bodiou Anitha Ajith Kumar Edoardo Massarelli Tessa van Haaften Mark J. Post Mark J. Post Panagiota Moutsatsou |
| author_facet | Vincent Bodiou Vincent Bodiou Anitha Ajith Kumar Edoardo Massarelli Tessa van Haaften Mark J. Post Mark J. Post Panagiota Moutsatsou |
| author_sort | Vincent Bodiou |
| collection | DOAJ |
| description | IntroductionTo 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.MethodsIn 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.Results and DiscussionOur 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 serum-free 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. |
| format | Article |
| id | doaj-art-c2e70c864a8c40679b641e12df92fa43 |
| institution | OA Journals |
| issn | 2296-4185 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj-art-c2e70c864a8c40679b641e12df92fa432025-08-20T01:47:25ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852024-11-011210.3389/fbioe.2024.14439141443914Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serumVincent Bodiou0Vincent Bodiou1Anitha Ajith Kumar2Edoardo Massarelli3Tessa van Haaften4Mark J. Post5Mark J. Post6Panagiota Moutsatsou7Mosa Meat BV, Maastricht, NetherlandsCARIM (The Cardiovascular Research Institute Maastricht), Faculty of Health, Medicine and Life Sciences, School for Cardiovascular Diseases, Maastricht University, Maastricht, NetherlandsMosa Meat BV, Maastricht, NetherlandsMosa Meat BV, Maastricht, NetherlandsMosa Meat BV, Maastricht, NetherlandsMosa Meat BV, Maastricht, NetherlandsCARIM (The Cardiovascular Research Institute Maastricht), Faculty of Health, Medicine and Life Sciences, School for Cardiovascular Diseases, Maastricht University, Maastricht, NetherlandsMosa Meat BV, Maastricht, NetherlandsIntroductionTo 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.MethodsIn 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.Results and DiscussionOur 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 serum-free 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.https://www.frontiersin.org/articles/10.3389/fbioe.2024.1443914/fullcultivated meatbovine myoblastsMicrobeadscell expansionproliferationbioprocessing |
| spellingShingle | Vincent Bodiou Vincent Bodiou Anitha Ajith Kumar Edoardo Massarelli Tessa van Haaften Mark J. Post Mark J. Post Panagiota Moutsatsou Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum Frontiers in Bioengineering and Biotechnology cultivated meat bovine myoblasts Microbeads cell expansion proliferation bioprocessing |
| title | Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum |
| title_full | Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum |
| title_fullStr | Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum |
| title_full_unstemmed | Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum |
| title_short | Attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum |
| title_sort | attachment promoting compounds significantly enhance cell proliferation and purity of bovine satellite cells grown on microcarriers in the absence of serum |
| topic | cultivated meat bovine myoblasts Microbeads cell expansion proliferation bioprocessing |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1443914/full |
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