Mesenchymal stroma cells (MSCs) have potential as an emerging cell therapy for treating many different diseases, but discovery of the practical sources of MSCs is needed for the large-scale clinical application of this therapy. This study was to identify MSCs in peritoneal dialysis (PD) effluents that were discarded after PD. The effluents were collected from patients who were on the dialysis for less than 1 month. Adherent cells from the effluents were isolated by incubation in serum-containing medium in plastic culture dishes. Cell surface markers were determined by a flow cytometric analysis, and the in vitro differentiation to chondrocytes, osteocytes or adipocytes was confirmed by staining with a specific dye. After four passages, these isolated cells displayed the typical morphology of mesenchymal cells in traditional 2-D cultures, and were grown to form spherical colonies in 3-D collagen cultures. Flow cytometric analysis revealed that the unsorted cells from all of seven patient samples showed robust expression of typical mesenchymal marker CD29, CD44, CD73, CD90 and CD166, and the absence of CD34, CD79a, CD105, CD271, SSEA-4, Stro-1 and HLA-DR. In differentiation assays, these cells were induced in vitro to chondrocytes, osteocytes or adipocytes. In conclusion, this preliminary study suggests the presence of MSCs in the "discarded" PD effluents. Further characterization of the phenotypes of these MSCs and evaluation of their therapeutic potential, particularly for the prevention of PD failure, are needed.

译文

间充质基质细胞(MSCs)作为治疗许多不同疾病的新兴细胞疗法具有潜力,但是该疗法的大规模临床应用需要发现MSCs的实际来源。这项研究是为了确定腹膜透析(PD)流出物中的MSC,这些MSC在PD后被丢弃。从透析时间少于1个月的患者中收集污水。通过在塑料培养皿中的含血清培养基中孵育,分离出流出物的粘附细胞。通过流式细胞术分析确定细胞表面标志物,并通过用特定染料染色确认体外分化为软骨细胞,骨细胞或脂肪细胞。经过四次传代后,这些分离的细胞在传统的2-D培养物中显示出间充质细胞的典型形态,并在3-D胶原培养物中生长形成球形菌落。流式细胞仪分析显示,来自所有七个患者样品的未分选细胞均表现出典型的间充质标记CD29,CD44,CD73,CD90和CD166的强表达,而不含CD34,CD79a,CD105,CD271,SSEA-4,Stro-1和HLA-DR。在分化测定中,这些细胞在体外被诱导成软骨细胞,骨细胞或脂肪细胞。总之,这项初步研究表明在“丢弃的” PD废水中存在MSC。需要进一步表征这些MSC的表型并评估其治疗潜力,尤其是对于预防PD衰竭。

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