Metcalf Garrett L, McClure Scott R, Hostetter Jesse M, Martinez Rudy F, Wang Chong
Department of Veterinary Clinical Sciences (Metcalf, McClure), Department of Pathology (Hostetter), Department of Diagnostics and Production Animal Medicine and Department of Statistics (Wang), Iowa State University, Ames, Iowa, USA; InGeneron Inc., 8205 El Rio, Houston, Texas, USA (Martinez).
Can J Vet Res. 2016 Oct;80(4):294-301.
Use of mesenchymal stem cells (MSCs) found in the stromal vascular fraction (SVF) of equine adipose tissue has promising applications for regenerative therapies. The most commonly used source of equine adipose tissue is the subcutaneous tailhead. The objective of this study was to compare 3 adipose depot sites in horses and determine the viability and cellular yield, capillary density, gene expression for selected markers, and colony-forming unit fibroblasts (CFU-Fs) in adipose tissue taken from these sites. Adipose tissue was excised from the area lateral to the tailhead, the inguinal region, and the small colon mesentery of 6 horses. Lipoaspirate was also collected from the area lateral to the tailhead. Stromal vascular fraction (SVF) was prepared in duplicate from the 3 different adipose tissue depots. The total nucleated and dead cell counts was determined manually using a hemocytometer and percent viability was calculated. Mass and volume of adipose were determined in order to calculate density and factor-VIII immunohistochemical staining was used to determine vascular density in the excisional adipose tissue samples from each horse. Quantitative polymerase chain reaction (qPCR) was used to quantify gene expression for selected cellular markers from each site. There were significant differences in viability, yield of nucleated cells/gram of adipose tissue, vascular density, gene expression, and CFU-Fs among adipose depots. Adipose from the mesentery yielded the highest number of nucleated cells/gram of tissue and the highest vascular density and percentage of CFU-Fs. In the horse, both the anatomical site of collection and the method of tissue collection significantly impact the yield and composition of cells in the SVF. Further study is needed to assess whether one adipose source is superior for harvesting mesenchymal stem cells (MSCs) and whether the differences among sources are clinically relevant for treatment of musculoskeletal injuries in horses.
在马脂肪组织的基质血管成分（SVF）中发现的间充质干细胞（MSC）的应用为再生疗法带来了广阔前景。马脂肪组织最常用的来源是皮下尾根部。本研究的目的是比较马的3个脂肪储存部位，并确定从这些部位获取的脂肪组织中的活力和细胞产量、毛细血管密度、选定标志物的基因表达以及集落形成单位成纤维细胞（CFU-F）。从6匹马的尾根外侧区域、腹股沟区和小结肠系膜处切除脂肪组织。还从尾根外侧区域收集了脂肪抽吸物。从3个不同的脂肪组织储存部位重复制备基质血管成分（SVF）。使用血细胞计数器手动确定总核细胞和死细胞数量，并计算活力百分比。测定脂肪的质量和体积以计算密度，并使用因子VIII免疫组织化学染色来确定每匹马切除的脂肪组织样本中的血管密度。定量聚合酶链反应（qPCR）用于量化每个部位选定细胞标志物的基因表达。脂肪储存部位之间在活力、每克脂肪组织的有核细胞产量、血管密度、基因表达和CFU-F方面存在显著差异。肠系膜脂肪产生的每克组织有核细胞数量最多，血管密度和CFU-F百分比最高。在马中，采集的解剖部位和组织采集方法均会显著影响SVF中细胞的产量和组成。需要进一步研究来评估一种脂肪来源在收获间充质干细胞（MSC）方面是否更具优势，以及不同来源之间的差异在临床上对治疗马的肌肉骨骼损伤是否具有相关性。
