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Methods and protocols2024; 7(2); 32; doi: 10.3390/mps7020032

Optimization of the Amplification of Equine Muscle-Derived Mesenchymal Stromal Cells in a Hollow-Fiber Bioreactor.

Abstract: The main causes of mortality in horses are the gastrointestinal pathologies associated with septic shock. Stem cells have shown, through systemic injection, a capacity to decrease inflammation and to regenerate injured tissue faster. Nevertheless, to achieve this rapid and total regeneration, systemic injections of 1 to 2 million cells per kilogram of body weight must be considered. Here, we demonstrate for the first time the feasibility and expansion capacity of equine muscle-derived mesenchymal stromal cells (mdMSCs) in a functionally closed, automated, perfusion-based, hollow-fiber bioreactor (HFBR) called the Quantum™ Cell Expansion System (Terumo Blood and Cell Technologies). This feature greatly increases the number of generated cells with a surface area of 1.7 m. The expansion of mdMSCs is very efficient in this bioreactor. The maximum expansion generated twenty times more cells than the initial seeding in nine days. The best returns were observed with an optimal seeding between 10 and 25 million mdMSCs, using the Bull's eye loading method and with a run duration between 7 and 10 days. Moreover, all the generated cells kept their stem properties: the ability to adhere to plastic and to differentiate into chondroblasts, osteoblasts and adipocytes. They also showed the expression of CD-44 and CD-90 markers, with a positive rate above 93%, while CD-45 and MHCII were non-expressed, with a positive rate below 0.5%. By capitalizing on the scalability, automation and 3D culture capabilities of the Quantum™, it is possible to generate large quantities of high-quality equine mdMSCs for gastrointestinal disorders and other clinical applications.
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Publication Date: 2024-04-02 PubMed ID: 38668139PubMed Central: PMC11054422DOI: 10.3390/mps7020032Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research studies the successful usage of a hollow-fiber bioreactor (a kind of laboratory equipment) to increase the number of equine muscle-derived mesenchymal stromal cells, a type of stem cell. This method could potentially improve the treatment of certain gastrointestinal conditions in horses.

Methodology

  • The researchers used a hollow-fiber bioreactor (HFBR), specifically the Quantum™ Cell Expansion System from Terumo Blood and Cell Technologies.
  • This system enables the increase in the number of cells generated and provides a surface area of 1.7 meters for cell growth.
  • The seeding (initial placement of cells to grow) of equine muscle-derived mesenchymal stromal cells (mdMSCs), a type of stem cell, was done optimally between 10 and 25 million mdMSCs. This was done using a technique known as the Bull’s eye loading method.
  • The procedures were conducted over a run duration of between 7 to 10 days.

Findings

  • The expansion of mdMSCs in the HFBR was highly successful, generating up to twenty times more cells than were initially seeded within nine days.
  • All the newly generated cells maintained their stem cell properties. These are the capacity to adhere to plastic and to differentiate, or transform, into other cells such as chondroblasts, osteoblasts and adipocytes.
  • The cells also showed expression of CD-44 and CD-90 markers (types of proteins characteristic of stem cells) at rates above 93%. They did not express CD-45 and MHCII, with rates below 0.5%.

Implications

  • This research suggests that the Quantum™ Cell Expansion System can be efficiently used to generate large quantities of high-quality equine mdMSCs.
  • This expanded production of stem cells could potentially be used in treating gastrointestinal disorders in horses. Given that such disorders are a common cause of horse mortality, this research provides hope for improved treatment options.
  • This system may also potentially be utilized for other clinical applications that require stem cells.

Cite This Article

APA
Duysens J, Graide H, Niesten A, Mouithys-Mickalad A, Ceusters J, Serteyn D. (2024). Optimization of the Amplification of Equine Muscle-Derived Mesenchymal Stromal Cells in a Hollow-Fiber Bioreactor. Methods Protoc, 7(2), 32. https://doi.org/10.3390/mps7020032

Publication

Methods and protocols
ISSN: 2409-9279
NlmUniqueID: 101720073
Country: Switzerland
Language: English
Volume: 7
Issue: 2
PII: 32

Researcher Affiliations

Duysens, Julien
  • Revatis SA, Rue de la Science 8, 6900 Marche-En-Famenne, Belgium.
  • Centre of Oxygen Research and Development (CORD), University of Liege, 4000 Liege, Belgium.
Graide, Hélène
  • Revatis SA, Rue de la Science 8, 6900 Marche-En-Famenne, Belgium.
Niesten, Ariane
  • Centre of Oxygen Research and Development (CORD), University of Liege, 4000 Liege, Belgium.
Mouithys-Mickalad, Ange
  • Centre of Oxygen Research and Development (CORD), University of Liege, 4000 Liege, Belgium.
Ceusters, Justine
  • Revatis SA, Rue de la Science 8, 6900 Marche-En-Famenne, Belgium.
  • Centre of Oxygen Research and Development (CORD), University of Liege, 4000 Liege, Belgium.
Serteyn, Didier
  • Revatis SA, Rue de la Science 8, 6900 Marche-En-Famenne, Belgium.
  • Centre of Oxygen Research and Development (CORD), University of Liege, 4000 Liege, Belgium.

Conflict of Interest Statement

D.S. and J.C. received a patent for mdMSCs (WO2015091210). D.S. is an administrator of the Revatis company, a provider of mdMSCs. H.G., J.D. and J.C. are employees at Revatis. The other authors have no conflicts of interest.

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Citations

This article has been cited 1 times.
  1. Xu Y, Liu X, Ahmad MA, Ao Q, Yu Y, Shao D, Yu T. Engineering cell-derived extracellular matrix for peripheral nerve regeneration. Mater Today Bio 2024 Aug;27:101125.
    doi: 10.1016/j.mtbio.2024.101125pubmed: 38979129google scholar: lookup
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