Chan Leo Li-Ying, Cohen Donald A, Kuksin Dmitry, Paradis Benjamin D, Qiu Jean
Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA,
J Fluoresc. 2014 Jul;24(4):983-9. doi: 10.1007/s10895-014-1388-x. Epub 2014 Apr 17.
In recent years, the lipoaspirate collected from adipose tissue has been seen as a valuable source of adipose-derived mesenchymal stem cells for autologous cellular therapy. For multiple applications, adipose-derived mesenchymal stem cells are isolated from the stromal vascular fraction (SVF) of adipose tissue. Because the fresh stromal vascular fraction typically contains a heterogeneous mixture of cells, determining cell concentration and viability is a crucial step in preparing fraction samples for downstream processing. Due to a large amount of cellular debris contained in the SVF sample, as well as counting irregularities standard manual counting can lead to inconsistent results. Advancements in imaging and optics technologies have significantly improved the image-based cytometric analysis method. In this work, we validated the use of fluorescence-based image cytometry for SVF concentration and viability measurement, by comparing to standard flow cytometry and manual hemocytometer. The concentration and viability of freshly collected canine SVF samples are analyzed, and the results highly correlated between all three methods, which validated the image cytometry method for canine SVF analysis, and potentially for SVF from other species.
近年来，从脂肪组织收集的脂肪抽吸物被视为用于自体细胞治疗的脂肪来源间充质干细胞的宝贵来源。对于多种应用，脂肪来源的间充质干细胞是从脂肪组织的基质血管部分（SVF）中分离出来的。由于新鲜的基质血管部分通常包含细胞的异质混合物，因此确定细胞浓度和活力是为下游处理准备部分样品的关键步骤。由于SVF样品中含有大量细胞碎片，以及计数不规则，标准的手动计数可能会导致结果不一致。成像和光学技术的进步显著改进了基于图像的细胞计数分析方法。在这项工作中，我们通过与标准流式细胞术和手动血细胞计数器比较，验证了基于荧光的图像细胞术用于SVF浓度和活力测量的有效性。分析了新鲜采集的犬类SVF样品的浓度和活力，所有三种方法的结果高度相关，这验证了图像细胞术用于犬类SVF分析的有效性，并可能适用于其他物种来源的SVF分析。
