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Culture and Immunomodulation of Equine Muscle-Derived Mesenchymal Stromal Cells: A Comparative Study of Innovative 2D Versus 3D Models Using Equine Platelet Lysate
by
J. Duysens
J. Duysens 1,2,*,†
,
H. Graide
H. Graide 1,†
,
A. Niesten
A. Niesten 2,
A. Mouithys-Mickalad
A. Mouithys-Mickalad
Ange Mouithys-Mickalad is a senior researcher at the Centre
of Oxygen Research and Development He [...]
Ange Mouithys-Mickalad is a senior researcher at the Centre
of Oxygen Research and Development (CORD). He obtained his PhD in medicinal
chemistry at the University of Lille II (France) in 1994. His area of interest
is the design, synthesis, and pharmacological study of new drugs susceptible to
be used as medicine for the treatment of schizophrenia and brain disorders.
These research works have been patented and published (French, UE, US, and Japanese
patents). Later on, he moved to Belgium (1995) and joined the team of Professor
Maurice Lamy, the former director of CORD, currently headed by Professor Didier
Serteyn. The current area of interest of the AMM is EPR applied to chemistry,
biology, and medicine with special emphasis on free radicals and oxidative
stress. He received in 2007 the title of Agrégé de Faculty of Medicine
(University of Liège).
2
,
G. Deby-Dupont
G. Deby-Dupont 2,
T. Franck
T. Franck 2
,
J. Ceusters
J. Ceusters 1,2 and
D. Serteyn
D. Serteyn
Didier Serteyn is a professor at the University of Liège and
CEO at Revatis SA. He graduated as a [...]
Didier Serteyn is a professor at the University of Liège and
CEO at Revatis SA. He graduated as a doctor of veterinary medicine from the
University of Liege in 1982 and obtained his PhD in 1989. As a full professor
of equine anesthesia, surgery, and intensive care, he is the head of the equine
clinic of the Faculty of Veterinary Medicine and was recently recognized as a
diplomate of the “European College of Veterinary Sports Medicine and
Rehabilitation”. His applied research activities have brought essential
knowledge in pathologies such as myopathy, laminitis, and colic and also
orthopedic developmental (osteochondrosis) and degenerative pathologies
(osteoarthritis). The themes of more fundamental research are related to
mitochondrial dysfunction, oxidative stress, free radical generation,
inflammation, and regenerative medicine. He is the promotor of 15 PhD programs
and the founder of two spin-off companies (BiopTis and RevaTis).
1,2
1
Revatis SA, Rue de la Science 8, 6900 Marche-En-Famenne, Belgium
2
Centre of Oxygen Research and Development (CORD), University of Liege, 4000 Liege, Belgium
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Cells 2024, 13(15), 1290; https://doi.org/10.3390/cells13151290 (registering DOI)
Submission received: 3 June 2024
/
Revised: 23 July 2024
/
Accepted: 29 July 2024
/
Published: 31 July 2024
Abstract
Muscle-derived mesenchymal stromal cells (mdMSCs) hold great promise in regenerative medicine due to their immunomodulatory properties, multipotent differentiation capacity and ease of collection. However, traditional in vitro expansion methods use fetal bovine serum (FBS) and have numerous limitations including ethical concerns, batch-to-batch variability, immunogenicity, xenogenic contamination and regulatory compliance issues. This study investigates the use of 10% equine platelet lysate (ePL) obtained by plasmapheresis as a substitute for FBS in the culture of mdMSCs in innovative 2D and 3D models. Using muscle microbiopsies as the primary cell source in both models showed promising results. Initial investigations indicated that small variations in heparin concentration in 2D cultures strongly influenced medium coagulation with an optimal proliferation observed at final heparin concentrations of 1.44 IU/mL. The two novel models investigated showed that expansion of mdMSCs is achievable. At the end of expansion, the 3D model revealed a higher total number of cells harvested (64.60 ± 5.32 million) compared to the 2D culture (57.20 ± 7.66 million). Trilineage differentiation assays confirmed the multipotency (osteoblasts, chondroblasts and adipocytes) of the mdMSCs generated in both models with no significant difference observed. Immunophenotyping confirmed the expression of the mesenchymal stem cell (MSC) markers CD-90 and CD-44, with low expression of CD-45 and MHCII markers for mdMSCs derived from the two models. The generated mdMSCs also had great immunomodulatory properties. Specific immunological extraction followed by enzymatic detection (SIEFED) analysis demonstrated that mdMSCs from both models inhibited myeloperoxidase (MPO) activity in a strong dose-dependent manner. Moreover, they were also able to reduce reactive oxygen species (ROS) activity, with mdMSCs from the 3D model showing significantly higher dose-dependent inhibition compared to the 2D model. These results highlighted for the first time the feasibility and efficacy of using 10% ePL for mdMSC expansion in novel 2D and 3D approaches and also that mdMSCs have strong immunomodulatory properties that can be exploited to advance the field of regenerative medicine and cell therapy instead of using FBS with all its drawbacks.
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MDPI and ACS Style
Duysens, J.; Graide, H.; Niesten, A.; Mouithys-Mickalad, A.; Deby-Dupont, G.; Franck, T.; Ceusters, J.; Serteyn, D.
Culture and Immunomodulation of Equine Muscle-Derived Mesenchymal Stromal Cells: A Comparative Study of Innovative 2D Versus 3D Models Using Equine Platelet Lysate. Cells 2024, 13, 1290.
https://doi.org/10.3390/cells13151290
AMA Style
Duysens J, Graide H, Niesten A, Mouithys-Mickalad A, Deby-Dupont G, Franck T, Ceusters J, Serteyn D.
Culture and Immunomodulation of Equine Muscle-Derived Mesenchymal Stromal Cells: A Comparative Study of Innovative 2D Versus 3D Models Using Equine Platelet Lysate. Cells. 2024; 13(15):1290.
https://doi.org/10.3390/cells13151290
Chicago/Turabian Style
Duysens, J., H. Graide, A. Niesten, A. Mouithys-Mickalad, G. Deby-Dupont, T. Franck, J. Ceusters, and D. Serteyn.
2024. "Culture and Immunomodulation of Equine Muscle-Derived Mesenchymal Stromal Cells: A Comparative Study of Innovative 2D Versus 3D Models Using Equine Platelet Lysate" Cells 13, no. 15: 1290.
https://doi.org/10.3390/cells13151290
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