Paebst Felicitas, Piehler Daniel, Brehm Walter, Heller Sandra, Schroeck Carmen, Tárnok Attila, Burk Janina
Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig, Germany; Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, University of Leipzig, Leipzig, Germany.
Cytometry A. 2014 Aug;85(8):678-87. doi: 10.1002/cyto.a.22491. Epub 2014 Jun 3.
Horses are an approved large animal model for therapies of the musculoskeletal system. Especially for tendon disease where cell-based therapy is commonly used in equine patients, the translation of achieved results to human medicine would be a great accomplishment. Immunophenotyping of equine mesenchymal stromal cells (MSCs) remains the last obstacle to meet the criteria of the International Society for Cellular Therapy (ISCT) definition of human MSCs. Therefore, the surface antigen expression of CD 29, CD 44, CD 73, CD 90, CD 105, CD 14, CD 34, CD 45, CD 79α, and MHC II in equine MSCs from adipose tissue, bone marrow, umbilical cord blood, umbilical cord tissue, and tendon tissue was analyzed using flow cytometry. Isolated cells from the different sources and donors varied in their expression pattern of MSC-defining antigens. In particular, CD 90 and 105 showed most heterogeneity. However, cells from all samples were robustly positive for CD 29 and CD 44, while being mostly negative for CD 73 and the exclusion markers CD 14, CD 34, CD 45, CD 79α and MHC II. Furthermore, it was evident that enzymes used for cell detachment after in vitro-culture affected the detection of antigen expression. These results emphasize the need of standardization of MSC isolation, culturing, and harvesting techniques. As the equine MSCs did not meet all criteria the ISCT defined for human MSCs, further investigations for a better characterization of the cell type should be conducted.
马是用于肌肉骨骼系统治疗的一种经认可的大型动物模型。特别是对于肌腱疾病，基于细胞的疗法在马科动物患者中普遍使用，将所取得的成果转化到人类医学上将是一项巨大成就。马间充质基质细胞（MSCs）的免疫表型分析仍然是满足国际细胞治疗协会（ISCT）对人类MSCs定义标准的最后一个障碍。因此，使用流式细胞术分析了来自脂肪组织、骨髓、脐带血、脐带组织和肌腱组织的马MSCs中CD 29、CD 44、CD 73、CD 90、CD 105、CD 14、CD 34、CD 45、CD 79α和MHC II的表面抗原表达。来自不同来源和供体的分离细胞在MSCs定义抗原的表达模式上存在差异。特别是，CD 90和105表现出最大的异质性。然而，所有样本中的细胞对CD 29和CD 44均呈强阳性，而对CD 73以及排除标记物CD 14、CD 34、CD 45、CD 79α和MHC II大多呈阴性。此外，很明显，体外培养后用于细胞分离的酶会影响抗原表达的检测。这些结果强调了MSCs分离、培养和收获技术标准化的必要性。由于马MSCs不符合ISCT为人类MSCs定义的所有标准，因此应进行进一步研究以更好地表征这种细胞类型。
