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猪脂肪间充质干细胞分离、鉴定、诱导及联合扩张器重建乳房模型的研究

Isolation, identification, induction of porcine adipose mesenchymal stem cells and research in breast reconstruction model with expander

作者: 喻兰雅  张晨芳  寇晓梅 
单位:                      广州中医药大学(广州510010)        
关键词:                     脂肪间充质干细胞;细胞分离;细胞诱导;皮肤扩张器;乳房重建          
分类号:
出版年·卷·期(页码):2014·33·4(369-373)
摘要:

目的 通过从脂肪组织中提取脂肪间充质干细胞,并进行培养鉴定、定向诱导,再联合皮肤扩张器在小型猪腹部皮下塑造乳房重建模型,探讨脂肪间充质干细胞用于乳房重建的可能性。方法 取小型猪颈背部皮下脂肪组织,用胶原酶消化法获得脂肪间充质干细胞,结合免疫荧光法,将所培养细胞用流式细胞技术分析细胞表面抗原,体外诱导其向脂肪细胞、成骨细胞分化,油红O染色、茜素红染色等方法检测细胞诱导分化结果。筛选脂肪间充质干细胞,将脂肪间充质干细胞联合扩张器注射于小型猪乳房模型观察。结果 流式细胞技术检测CD34、CD45阴性表达,CD44、CD105阳性表达,油红O染色可见细胞内有脂滴形成,茜素红染色发现细胞出现钙结节。动物模型发现与对照组相比较,注入干细胞组局部细胞数目增多。结论 猪脂肪间充质干细胞可在体外培养,并可定向诱导成脂肪细胞、成骨细胞,在动物体内可向局部组织分化,引起局部细胞数目增多。

Objective To investigate the potential of adipose derived stem cells in reconstruction, we cultured and induced the adipose mesenchymal stem cells which were obtained from porcine adipose tissue, then set up a model of breast reconstruction with the skin expander in minipig’s subcutaneous abdominal adipose tissue. Methods Porcine adipose derived stem cells were obtained from subcutaneous adipose tissue of minipig with collagenase digestion, identified by flow cytometric analysis and induced to adopogenic and osteogenic differentiation in vitro. Results The stem cells were negative for CD34 and CD45, positive for CD44 and CD105. The cells were positive for oil red O staining and alizarin red staining. The local cells were much more in experimental group than in control group. Conclusions Porcine adipose derived stem cells can be cultured in vitro and induced adopogenic and osteogenic differentiation. The cells can be used in breast reconstruction model in vivo.

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