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尿液来源的细胞是泌尿外科组织重建的潜在细胞来源。

Urine derived cells are a potential source for urological tissue reconstruction.

作者信息

Zhang Yuanyuan, McNeill Elena, Tian Hong, Soker Shay, Andersson Karl-Erik, Yoo James J, Atala Anthony

机构信息

Institute for Regenerative Medicine, School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27157, USA.

出版信息

J Urol. 2008 Nov;180(5):2226-33. doi: 10.1016/j.juro.2008.07.023. Epub 2008 Sep 20.

Abstract

PURPOSE

Contemporary approaches to tissue engineering and cell therapy for urinary tract reconstruction require invasive tissue biopsies to obtain autologous cells. However, these procedures are associated with potential complications. We determined whether the cells present in urine have characteristics of normal bladder cells and investigated their potential uses for urological reconstructive procedures.

MATERIALS AND METHODS

A total of 55 urine samples were collected from 15 healthy individuals and 8 patients with vesicoureteral reflux. Urine derived cells were isolated, expanded and tested for progenitor and differentiated cell specific markers using flow cytometry, immunofluorescence and Western immunoblotting. The chromosomal stability of cultured urine derived cells was determined by karyotype analysis.

RESULTS

Clones were successfully established from primary cultures of urine derived cells. Isolated cells showed 3 phenotypes, including fully differentiated, differentiating and progenitor-like cells. Some urine derived cells stained positive for the surface markers c-Kit, SSEA4, CD105, CD73, CD91, CD133 and CD44. Two to 7 cells per 100 ml urine were multipoint progenitors that could expand extensively in culture. Single progenitor cells had the ability to differentiate into the cell lineages expressing urothelial, smooth muscle, endothelial and interstitial cell markers. The expression of lineage markers was characterized by Western blot and immunofluorescence analysis. Urine derived cells also maintained a normal karyotype after serial culture.

CONCLUSIONS

A subpopulation of cells isolated from urine had progenitor cell features and the potential to differentiate into several bladder cell lineages. Urine derived cells could serve as an alternative cell source for urinary tract tissue engineering and reconstruction.

摘要

目的

当代用于尿路重建的组织工程和细胞治疗方法需要进行侵入性组织活检以获取自体细胞。然而,这些操作存在潜在并发症。我们确定尿液中存在的细胞是否具有正常膀胱细胞的特征,并研究它们在泌尿外科重建手术中的潜在用途。

材料和方法

从15名健康个体和8名膀胱输尿管反流患者中总共收集了55份尿液样本。分离、扩增尿液来源的细胞,并使用流式细胞术、免疫荧光和Western免疫印迹法检测祖细胞和分化细胞特异性标志物。通过核型分析确定培养的尿液来源细胞的染色体稳定性。

结果

成功从尿液来源细胞的原代培养物中建立了克隆。分离的细胞表现出3种表型,包括完全分化、正在分化和祖细胞样细胞。一些尿液来源的细胞表面标志物c-Kit、SSEA4、CD105、CD73、CD91、CD133和CD44染色呈阳性。每100 ml尿液中有2至7个细胞是多能祖细胞,可在培养中广泛扩增。单个祖细胞有能力分化为表达尿路上皮、平滑肌、内皮和间质细胞标志物的细胞谱系。通过Western印迹和免疫荧光分析对谱系标志物的表达进行了表征。尿液来源的细胞在连续培养后也保持正常核型。

结论

从尿液中分离出的细胞亚群具有祖细胞特征,并有分化为几种膀胱细胞谱系的潜力。尿液来源的细胞可作为尿路组织工程和重建的替代细胞来源。

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