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用于大鼠膀胱再生的毛囊干细胞:初步结果

Hair stem cells for bladder regeneration in rats: preliminary results.

作者信息

Drewa T, Joachimiak R, Kaznica A, Sarafian V, Pokrywczynska M

机构信息

Department of Tissue Engineering, Nicolaus Copernicus University, ul. Karlowicza 24, 85-092 Bydgoszcz, Poland.

出版信息

Transplant Proc. 2009 Dec;41(10):4345-51. doi: 10.1016/j.transproceed.2009.08.059.

Abstract

BACKGROUND

A variety of tissue engineering techniques are currently under development or investigation for bladder augmentation, but so far no approach is clearly superior. The aim of this study was to compare the suitability for cystoplasty augmentation in rats of in vivo implanted acellular bladder matrices (BAM) previously seeded with hair follicle stem cells and that of matrices implanted without the cells.

MATERIALS AND METHODS

The rat hair follicle stem cell line was positive for CD34, p63, and Ki-67. 1 x 10(6) cells from 34 to 40 passages seeded onto nine BAM scaffolds were cultured for one week. Nine other scaffolds were left unseeded. Scaffolds were grafted into a surgically created defect within the anterior bladder wall: nine rats with acellular matrices and nine with cell-seeded BAM. Rats observed for six months were killed in monthly intervals. We performed gross examination, X-ray cystography, and hematoxylin-eosin, cytokine (CK)-7, CK-20, myoglobin, and desmin staining of the excised bladders.

RESULTS

Minimal adhesions were observed and urinary leakage was noted in one case. Two animals died in the acellular group. Rats developed stone disease in bladders reconstructed with acellular BAM. Bladder capacity was similar, but the shape was regular and characteristically oval only in bladders grafted with cell-seeded BAM. Muscle layers in the apical parts of the reconstructed bladder walls were extremely thin in the cases of acellular grafts and thicker in bladders reconstructed with cell-seeded grafts. Muscle layer regeneration was better in the cell-seeded group. Urothelium regenerated in all animals.

CONCLUSIONS

We have shown that hair follicle stem cells may be used for rat bladder wall regeneration.

摘要

背景

目前有多种组织工程技术正在开发或研究用于膀胱扩大术,但到目前为止,尚无一种方法明显更具优势。本研究的目的是比较预先接种毛囊干细胞的体内植入脱细胞膀胱基质(BAM)与未接种细胞的基质在大鼠膀胱扩大成形术中的适用性。

材料与方法

大鼠毛囊干细胞系CD34、p63和Ki-67呈阳性。将传代34至40代的1×10⁶个细胞接种到9个BAM支架上培养1周。另外9个支架未接种细胞。将支架移植到手术造成的膀胱前壁缺损处:9只大鼠植入脱细胞基质,9只大鼠植入接种细胞的BAM。对观察6个月的大鼠每隔1个月处死一批。我们对切除的膀胱进行大体检查、膀胱造影X线检查以及苏木精-伊红染色、细胞角蛋白(CK)-7、CK-20、肌红蛋白和结蛋白染色。

结果

观察到粘连极少,1例出现尿漏。脱细胞组有2只动物死亡。用脱细胞BAM重建膀胱的大鼠出现结石病。膀胱容量相似,但仅在接种细胞的BAM移植的膀胱中形状规则且呈典型椭圆形。脱细胞移植的重建膀胱壁顶部的肌肉层极薄,而接种细胞移植重建的膀胱中肌肉层较厚。接种细胞组的肌肉层再生更好。所有动物的尿路上皮均再生。

结论

我们已经表明毛囊干细胞可用于大鼠膀胱壁再生。

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