Production of extracellular vesicles from equine embryo-derived mesenchymal stromal cells.
Abstract: Mesenchymal stromal cell (MSC)-derived extracellular vesicles (EVs) have shown promise as off-the-shelf therapeutics; however, producing them in sufficient quantities can be challenging. In this study, MSCs were isolated from preimplantation equine embryos and used to produce EVs in two commercially available bioreactor designs. Mesenchymal stromal cells (MSC) have recently been explored for their potential use as therapeutics in human and veterinary medicine applications, such as the treatment of endometrial inflammation and infertility. Allogeneic MSC-derived extracellular vesicles (EVs) may also provide therapeutic benefits with advantage of being an 'off-the-shelf' solution, provided they can be produced in large enough quantities, without contamination from bovine EVs contained in fetal bovine serum that is a common component of cell culture media. Toward this aim, we demonstrated the successful isolation and characterization of equine MSCs from preimplantation embryos. We also demonstrate that many of these lines can be propagated long-term in culture while retaining their differentiation potential and conducted a head-to-head comparison of two bioreactor systems for scalable EV production including in serum-free conditions. Based on our findings, the CELLine AD 1000 flasks enabled higher cell density cultures and significantly more EV production than the FiberCell system or conventional culture flasks. These findings will enable future isolation of equine MSCs and the scalable culture of their EVs for a wide range of applications in this rapidly growing field.
Publication Date: 2022-08-25 PubMed ID: 35938796DOI: 10.1530/REP-22-0215Google Scholar: Lookup
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- Journal Article
Summary
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The research article studies the isolation of mesenchymal stromal cells (MSCs) from preimplantation equine embryos for the production of extracellular vesicles (EVs) that can serve as a viable treatment option in human and veterinary medicine. Specifically, the paper validates the possibility of using these equine derived EVs as ‘off-the-shelf’ therapeutics, considering their scalable production in two different bioreactor models.
Objective of the Study
- The primary goal of this research is to explore the potential of mesenchymal stromal cells (MSCs) for their therapeutic applications, specifically through the production of extracellular vesicles (EVs), derived from preimplantation equine embryos.
- The researchers aim to establish a robust, scalable method for the production of these EVs. The idea is to overcome the challenges involved in mass production and develop these vesicles as off-the-shelf therapeutics, ideally void of bovine EV contamination resulting from the use of fetal bovine serum in cell culture media.
Process and Results
- The researchers successfully isolated and characterized equine MSCs from preimplantation embryos. They then demonstrated that many of these isolated cell lines could be propagated long-term in cultures while still retaining their differentiation potential.
- To optimize the scalable production of EVs, two different commercially available bioreactor designs were evaluated: the CELLine AD 1000 flasks and the FiberCell system.
- Findings revealed that the CELLine AD 1000 flasks enabled higher cell density cultures, resulting in significantly more EV production than the FiberCell system or conventional culture flasks.
Conclusion and Future Implications
- This research plays a significant role in paving the way for future isolation of equine MSCs and their large-scale EV production, thereby extending their potential application in therapies across various medical fields.
- The findings highlight the potential of equine embryo-derived MSCs and their EVs as promising off-the-shelf solutions in human and veterinary medicine.
- Moreover, it broadens the scope for further studies to enhance the use of these equine MSC-derived EVs in treating various conditions such as endometrial inflammation and infertility.
Cite This Article
APA
Tasma Z, Hou W, Damani T, Seddon K, Kang M, Ge Y, Hanlon D, Hollinshead F, Hisey CL, Chamley LW.
(2022).
Production of extracellular vesicles from equine embryo-derived mesenchymal stromal cells.
Reproduction, 164(4), 143-154.
https://doi.org/10.1530/REP-22-0215 Publication
Researcher Affiliations
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Animal Reproduction and Biotechnology Laboratory, Department of Clinical Sciences, Colorado State University, Fort Collins, Colorado, USA.
- Animal Reproduction and Biotechnology Laboratory, Department of Clinical Sciences, Colorado State University, Fort Collins, Colorado, USA.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Hub for Extracellular Vesicle Investigations, The University of Auckland, Auckland, New Zealand.
- Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio, USA.
- Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand.
- Hub for Extracellular Vesicle Investigations, The University of Auckland, Auckland, New Zealand.
MeSH Terms
- Animals
- Cell Differentiation
- Embryo, Mammalian
- Extracellular Vesicles / metabolism
- Horses
- Humans
- Mesenchymal Stem Cells / metabolism
Citations
This article has been cited 7 times.- Saadeldin IM, Ehab S, Assiri AM. The Potential Roles of Extracellular Vesicles in Wild Animals' Reproduction. Adv Exp Med Biol 2026;16:209-223.
- Hisey CL, Rima XY, Doon-Ralls J, Nagaraj CK, Mayone S, Nguyen KT, Wiggins S, Dorayappan KDP, Huang X, Hade MD, Selvendiran K, Higginbotham JN, Tutanov O, Franklin JL, Coffey RJ, Wood D, Hu C, Patel DS, Magaña SM, Palmer AF, Hansford D, Reátegui E. Light-induced extracellular vesicle and particle adsorption. Nat Methods 2025 Dec;22(12):2609-2621.
- Barrachina L, Ivanovska A, Eslami Arshaghi T, O'Brien A, Cequier A, Murphy M, Hollinshead F, Rodellar C, Barry F. Generation of equine induced pluripotent stem cells from cells of embryonic, perinatal and adult tissues. Stem Cell Res Ther 2025 Oct 8;16(1):547.
- DePalma TJ, Hisey CL, Hughes K, Fraas D, Tawfik M, Scharenberg J, Wiggins S, Nguyen KT, Hansford DJ, Reátegui E, Skardal A. Tuning a bioengineered hydrogel for studying astrocyte reactivity in glioblastoma. Acta Biomater 2024 Nov;189:155-167.
- Kazemzadeh M, Martinez-Calderon M, Otupiri R, Artuyants A, Lowe M, Ning X, Reategui E, Schultz ZD, Xu W, Blenkiron C, Chamley LW, Broderick NGR, Hisey CL. Deep autoencoder as an interpretable tool for Raman spectroscopy investigation of chemical and extracellular vesicle mixtures. Biomed Opt Express 2024 Jul 1;15(7):4220-4236.
- Hisey CL, Artuyants A, Guo G, Chang V, Reshef G, Middleditch M, Jacob B, Chamley LW, Blenkiron C. Investigating the consistency of extracellular vesicle production from breast cancer subtypes using CELLine adherent bioreactors. J Extracell Biol 2022 Sep;1(9):e60.
- Gaesser AM, Usimaki AIJ, Barot DA, Linardi RL, Molugu S, Musante L, Ortved KF. Equine mesenchymal stem cell-derived extracellular vesicle productivity but not overall yield is improved via 3-D culture with chemically defined media. J Am Vet Med Assoc 2024 Jun 1;262(S1):S97-S108.
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