Reconstitution of Experimental Neurogenic Bladder Dysfunction Using Skeletal Muscle-Derived Multipotent Stem Cells
Reconstitution of Experimental Neurogenic Bladder Dysfunction Using Skeletal Muscle-Derived Multipotent Stem Cells
复制标题
DOI：
10.1097/tp.0b013e3181d45a7f
复制
发表时间：
2010-05-15
期刊：
影响因子：
6.2
通讯作者：
Terachi, Toshiro
中科院分区：
文献类型：
--
作者：
Nitta, Masahiro;Tamaki, Tetsuro;Terachi, Toshiro
背景资料。术后神经源性膀胱功能障碍是宫颈癌根治性子宫切除术的主要并发症，其主要原因是伴侧血管对盆丛膀胱支的不可避免的损伤。因此，我们尝试使用骨骼肌来源的多潜能干细胞重建受损的BBPP和血管，这些干细胞显示出血管、肌肉和周围神经系统的同步重建能力。在戊巴比妥钠麻醉下，测定电刺激BBPP所产生的膀胱内压作为膀胱功能。然后切断带血管的BBPP远端(实验性神经源性膀胱模型)。分别于移植前、移植后即刻、移植后4周进行功能恢复测量。从右侧比目鱼肌和腓肠肌经酶消化和细胞分选后，获得CD34(+)/45(-)(Sk-34)和CD34(-)/45(-)(Sk-dN)干细胞。将悬浮细胞自体移植到损伤区周围，对照组仅移植培养液和CD45(+)细胞。将从绿色荧光蛋白转基因小鼠肌肉中获得的干细胞移植到裸鼠模型中，进行免疫组织化学和免疫电子显微镜观察，以确定移植细胞的形态贡献。术后4周，移植组功能恢复率(接近80%)明显高于对照组(分别为28%和24%)。移植细胞分化为雪旺细胞、神经周围细胞、血管平滑肌细胞、周细胞和膀胱周围成纤维细胞后，可整合到受损的周围神经和血管内。多潜能Sk-34和Sk-dN细胞移植对于修复受损的BBPP具有潜在的应用价值。
Background. Postoperative neurogenic bladder dysfunction is a major complication of radical hysterectomy for cervical cancer and is mainly caused by unavoidable damage to the bladder branch of the pelvic plexus (BBPP) associated with colateral blood vessels. Thus, we attempted to reconstitute disrupted BBPP and blood vessels using skeletal muscle-derived multipotent stem cells that show synchronized reconstitution capacity of vascular, muscular, and peripheral nervous systems.Methods. Under pentobarbital anesthesia, intravesical pressure by electrical stimulation of BBPP was measured as bladder function. The distal portion of BBPP with blood vessels was then cut unilaterally (experimental neurogenic bladder model). Measurements were performed before, immediately after, and at 4 weeks after transplantation as functional recovery. Stem cells were obtained from the right soleus and gastrocnemius muscles after enzymatic digestion and cell sorting as CD34(+)/45(-) (Sk-34) and CD34(-)/45(-) (Sk-DN). Suspended cells were autografted around the damaged region, whereas medium alone and CD45(+) cells were transplanted as control groups. To determine the morphological contribution of the transplanted cells, stem cells obtained from green fluorescent protein transgenic mouse muscles were transplanted into a nude rat model and were examined by immunohistochemistry and immuno-electron microscopy.Results. At 4 weeks after surgery, the transplantation group showed significantly higher functional recovery (similar to 80%) than the two controls (similar to 28% and 24%). The transplanted cells showed an incorporation into the damaged peripheral nerves and blood vessels after differentiation into Schwann cells, perineurial cells, vascular smooth muscle cells, pericytes, and fibroblasts around the bladder.Conclusion. Transplantation of multipotent Sk-34 and Sk-DN cells is potentially useful for the reconstitution of damaged BBPP.