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.
Reproduction. 2022 Aug 25;164(4):143-154. doi: 10.1530/REP-22-0215. Print 2022 Oct 1.
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.
间充质基质细胞 (MSC) 最近在人类和兽医医学应用中被探索用于治疗，例如治疗子宫内膜炎和不孕。同种异体 MSC 衍生的细胞外囊泡 (EV) 也可能具有治疗益处，因为它们是一种“现成的”解决方案，前提是它们可以大量生产，而不会受到胎牛血清中牛 EV 的污染，胎牛血清是细胞培养基的常见成分。为此，我们成功地从胚胎中分离和鉴定了马 MSC。我们还证明，许多这些系可以在长期培养中繁殖，同时保持其分化潜能，并对两种用于可扩展 EV 生产的生物反应器系统进行了头对头比较，包括在无血清条件下。基于我们的发现，CELLine AD 1000 瓶使细胞密度培养更高，并显著增加了 EV 的产生，而 FiberCell 系统或传统培养瓶则不然。这些发现将使未来能够在这个快速发展的领域中分离马 MSC 并大规模培养其 EV，以满足广泛的应用需求。
