Rivera Orsini Michael A, Ozmen Emine Berfu, Miles Alyssa, Newby Steven D, Springer Nora, Millis Darryl, Dhar Madhu
Regenerative Medicine and Tissue Engineering, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA.
Genome Science and Technology, University of Tennessee Knoxville, Knoxville, TN 37996, USA.
Animals (Basel). 2024 Oct 15;14(20):2974. doi: 10.3390/ani14202974.
In allogeneic MSC implantation, the cells are isolated from a donor different from the recipient. When tested, allogeneic MSCs have several advantages over autologous ones: faster cell growth, sufficient cell concentration, and readily available cells for clinics. To ensure the safe and efficient use of allogeneic MSCs in clinics, the MSCs need to be first tested in vitro. With this study, we paved the way by addressing the in vitro aspects of canine adipose-derived MSCs, considering the limited studies on the clinical use of canine cells. We isolated cAD-MSCs from canine falciform ligament fat and evaluated their viability and proliferation using an MTS assay. Then, we characterized the MSC-specific antigens using immunophenotyping and immunofluorescence and demonstrated their potential for in vitro differentiation. Moreover, we established shipping and cryobanking procedures to lead the study to become an off-the-shelf therapy. During expansion, the cells demonstrated a linear increase in cell numbers, confirming their proliferation quantitatively. The cells showed viability before and after cryopreservation, demonstrating that cell viability can be preserved. From a clinical perspective, the established shipping conditions demonstrated that the cells retain their viability for up to 48 h. This study lays the groundwork for the potential use of allogeneic cAD-MSCs in clinical applications.
在异体间充质干细胞植入中，细胞是从与受体不同的供体中分离出来的。经测试，异体间充质干细胞相对于自体间充质干细胞具有几个优势：细胞生长更快、细胞浓度充足以及临床可用细胞易于获取。为确保异体间充质干细胞在临床中的安全有效使用，需要首先在体外对其进行测试。通过这项研究，鉴于对犬类细胞临床应用的研究有限，我们通过解决犬类脂肪来源间充质干细胞的体外相关问题，为后续研究铺平了道路。我们从犬类镰状韧带脂肪中分离出犬类脂肪来源间充质干细胞（cAD-MSCs），并使用MTS检测法评估其活力和增殖情况。然后，我们通过免疫表型分析和免疫荧光对间充质干细胞特异性抗原进行了表征，并证明了它们的体外分化潜力。此外，我们建立了运输和冷冻保存程序，以使该研究发展成为一种现成可用的治疗方法。在扩增过程中，细胞数量呈线性增加，从数量上证实了它们的增殖。细胞在冷冻保存前后均显示出活力，表明细胞活力可以得到保留。从临床角度来看，所建立的运输条件表明细胞在长达48小时内仍保持其活力。这项研究为异体cAD-MSCs在临床应用中的潜在用途奠定了基础。
