Mature, Adipocyte Derived, Dedifferentiated Fat Cells Can Differentiate Into Smooth Muscle-Like Cells and Contribute to Bladder Tissue Regeneration
Mature, Adipocyte Derived, Dedifferentiated Fat Cells Can Differentiate Into Smooth Muscle-Like Cells and Contribute to Bladder Tissue Regeneration
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DOI：
10.1016/j.juro.2009.02.103
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发表时间：
2009-07-01
影响因子：
6.6
通讯作者：
Mugishima, Hideo
中科院分区：
文献类型：
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作者：
Sakuma, Takahiro;Matsumoto, Taro;Mugishima, Hideo
目的：我们最近报道了成熟的、脂肪细胞衍生的、去分化的脂肪细胞具有高增殖活性和多谱系分化潜力。在目前的研究中，我们研究了这些细胞是否可以分化成平滑肌细胞谱系，并在小鼠膀胱损伤模型中促进膀胱组织再生。材料与方法：将人脂肪细胞来源的去分化脂肪细胞在平滑肌细胞分化条件下培养1周，并进行a-平滑肌肌动蛋白免疫染色。实时逆转录-聚合酶链反应检测脂肪细胞分化平滑肌细胞标记基因的表达。将绿色荧光蛋白标记的去分化脂肪细胞注射到低温损伤小鼠膀胱壁上。移植后14天和30天，用免疫组织化学方法检测脂肪细胞再生平滑肌组织的能力。结果：免疫组化分析显示，在含5%胎牛血清和5 ng/ml转化生长因子- β 1的最佳培养条件下，50%以上的脂肪细胞成功分化为α -平滑肌肌动蛋白阳性细胞。实时逆转录聚合酶链反应显示，在分化培养的第1周，去分化脂肪细胞中SM22 α、α -平滑肌肌动蛋白和平滑肌-肌球蛋白重链的表达增加。在移植后14天和30天小鼠膀胱壁注射部位观察到表达a-平滑肌肌动蛋白和绿色荧光蛋白的细胞。脂肪细胞注射小鼠损伤膀胱组织α -平滑肌肌动蛋白阳性区明显大于生理盐水注射对照组。结论：去分化脂肪细胞可分化为平滑肌细胞系，促进膀胱平滑肌组织再生。
Purpose: We recently reported that mature, adipocyte derived, dedifferentiated fat cells show high proliferative activity and multilineage differentiation potential. In the current study we investigated whether such cells could differentiate into a smooth muscle cell lineage and contribute to bladder tissue regeneration in a mouse bladder injury model.Materials and Methods: Human adipocyte derived dedifferentiated fat cells were cultured for 1 week under conditions favorable for smooth muscle cell differentiation and immunostained for a-smooth muscle actin. The expression of smooth muscle cell marker genes for differentiating dedifferentiated fat cells was measured by real-time reverse transcription-polymerase chain reaction. Green fluorescence protein labeled dedifferentiated fat cells were injected into cryo-injured bladder walls in mice. The ability of the fat cells to regenerate smooth muscle tissue was examined immunohistochemically 14 and 30 days after transplantation.Results: Immunohistochemical analysis revealed that more than 50% of the fat cells were successfully differentiated into alpha-smooth muscle actin positive cells under the optimum culture condition of a medium containing 5% fetal bovine serum and 5 ng/ml transforming growth factor-beta 1. Real-time reverse transcription-polymerase chain reaction revealed increased expression of SM22 alpha, alpha-smooth muscle actin and smooth muscle-myosin heavy chain in dedifferentiated fat cells during week 1 of differentiation culture. Cells expressing a-smooth muscle actin plus green fluorescence protein were observed at the bladder wall injection sites in mice 14 and 30 days after transplantation. alpha-Smooth muscle actin positive areas in injured bladder tissue in mice with fat cell injection were significantly larger than those in saline injected control mice.Conclusions: These findings suggest that dedifferentiated fat cells can differentiate into smooth muscle cell lineages and contribute to the regeneration of bladder smooth muscle tissue.