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Zoe Tasma Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand

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Weilin Hou Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand

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Tanvi Damani Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand

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Kathleen Seddon Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand

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Matthew Kang Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand

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Yi Ge Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand

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David Hanlon Animal Reproduction and Biotechnology Laboratory, Department of Clinical Sciences, Colorado State University, Fort Collins, Colorado, USA

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Fiona Hollinshead Animal Reproduction and Biotechnology Laboratory, Department of Clinical Sciences, Colorado State University, Fort Collins, Colorado, USA

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Colin L Hisey 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

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Lawrence W Chamley Department of Obstetrics and Gynaecology, The University of Auckland, Auckland, New Zealand
Hub for Extracellular Vesicle Investigations, The University of Auckland, Auckland, New Zealand

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In brief

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.

Abstract

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.

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