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A Simplified 3D-Plasma Culture Method for Generating Minimally Manipulated Autologous Equine Muscle-Derived Progenitor Cells.
Abstract: Musculoskeletal pathologies present challenges in athletic horses, often leading to functional impairment. The slow or limited regenerative capacity of bone, joint, and tendon/ligament injuries, coupled with the limitations of conventional treatments, highlights the need for innovative therapies such as ortho-biologics and mesenchymal stem/stroma cells. Traditional 2D cell culture systems with fetal bovine serum (FBS) fail to replicate the complexity of the in vivo environment, whereas 3D cultures more accurately mimic native tissue architecture and cell-cell interactions. This study describes a novel method for isolating muscle-derived progenitor cells in a 3D environment using an autologous plasma-based gel and an innovative cell retrieval solution. The cultured cells exhibit immunomodulatory effects on T lymphocytes, trilineage differentiation potential, and immunophenotypic characteristics consistent with conventional mesenchymal stem/stromal cells. This streamlined 3D culture technique offers a promising platform for generating minimally manipulated autologous cell products tailored for equine regenerative medicine. Key features • Development of a simplified autologous 3D-plasma-based culture method, eliminating the need for fetal bovine serum. • Fully autologous approach offering safer clinical potential for equine regenerative medicine. • Easy and time-efficient method suitable for researchers familiar with basic cell culture techniques.
©Copyright : © 2025 The Authors; This is an open access article under the CC BY-NC license.
Publication Date: 2025-12-05 PubMed ID: 41384097PubMed Central: PMC12689254DOI: 10.21769/BioProtoc.5526Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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Overview
- This study presents a new, simplified method for growing muscle-derived progenitor cells from horses using their own plasma in a three-dimensional culture system.
- The method avoids fetal bovine serum, better mimics the natural tissue environment, and produces cells with properties suitable for regenerative therapies in horses.
Background and Motivation
- Musculoskeletal injuries in athletic horses, such as those affecting bones, joints, tendons, and ligaments, often heal slowly or incompletely, leading to long-term functional problems.
- Conventional treatments have limitations, necessitating new therapeutic options like ortho-biologics and mesenchymal stem/stromal cell (MSC) therapies.
- Traditional cell culture methods use two-dimensional (2D) systems with fetal bovine serum (FBS), which do not replicate the complex three-dimensional (3D) environment of living tissues, possibly affecting cell behavior and therapy outcomes.
- 3D culture systems better mimic the natural cellular environment by preserving cell-cell and cell-matrix interactions, potentially improving cell functionality for regenerative medicine.
Novel Method Developed
- The researchers developed a simplified 3D culture method using a gel made from the horse’s own plasma, eliminating the need for FBS and thus reducing risks of immune reactions and contamination.
- This method is fully autologous, meaning all components come from the same horse, enhancing the safety profile for clinical use.
- The technique includes an innovative cell retrieval solution that allows for efficient extraction of cells grown in the 3D plasma gel without damaging their properties.
- The entire culture process is designed to be easy to perform and time-efficient, making it accessible for laboratories familiar with basic cell culture techniques.
Characteristics of Cultured Cells
- The muscle-derived progenitor cells grown in this 3D plasma-based system exhibit immunomodulatory effects on T lymphocytes, indicating potential to regulate immune responses beneficially.
- These cells demonstrate trilineage differentiation potential, meaning they can develop into three different types of cells (such as bone, cartilage, and fat), a hallmark of mesenchymal stem/stromal cells.
- The immunophenotypic profile (surface markers and other characteristics) of these cultured cells aligns with conventional MSCs, confirming their identity and potential utility.
Clinical and Research Implications
- This method paves the way for producing minimally manipulated autologous cell products tailored for equine regenerative medicine applications.
- Because it avoids animal serum products and uses the horse’s own biological materials, it may improve safety and reduce regulatory hurdles for clinical use in horses.
- The approach also offers researchers a practical platform to generate muscle progenitor cells in a more physiologically relevant 3D environment, potentially improving experimental models.
- Ultimately, this work could contribute to improved therapies for musculoskeletal injuries in athletic horses, enhancing recovery and performance.
Cite This Article
APA
Graide H, Duysens J, Frank T, Mouithys-Mickalad A, Niesten A, Sandersen C, Ceusters J, Serteyn D.
(2025).
A Simplified 3D-Plasma Culture Method for Generating Minimally Manipulated Autologous Equine Muscle-Derived Progenitor Cells.
Bio Protoc, 15(23), e5526.
https://doi.org/10.21769/BioProtoc.5526 Publication
Researcher Affiliations
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Department of equine clinical sciences, University of Liège, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Department of equine clinical sciences, University of Liège, Liège, Belgium.
- Center for Oxygen Research and Development, University of Liege, Liège, Belgium.
- Department of equine clinical sciences, University of Liège, Liège, Belgium.
Conflict of Interest Statement
Competing interestsD. Serteyn and J. Ceusters are the co-inventors of a patent related to muscle-derived stem cells. This patent is licensed by the University of Liège to Revatis, a Spin-Off company where D. Serteyn and J. Ceusters serve as scientific advisors. The other co-authors declare no conflicts of interest.
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