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小鼠骨髓来源干细胞在皮肤创伤修复中的募集作用观察
Recruitment effect of bone marrow derived stem cells (BMSCs) on the repair of dermal wound
【摘要】 目的观察小鼠骨髓来源干细胞在皮肤创伤修复中的动员及募集作用。方法提取雄性C57小鼠骨髓成体干细胞,采用差异性贴壁法分离纯化骨髓成体干细胞。取第三代细胞进行成骨、成脂诱导。同时另取所培养的第三代细胞,用BrDU标记,胰酶消化后,调整细胞浓度为1×108/ml,经尾静脉注射到5.0Gy放射性损伤后的雌性小鼠体内(n=6),建立骨髓移植嵌合模型。3d后建立皮肤缺损模型,于创伤7d后取创面组织,观察是否募集有BrDU阳性细胞;并于皮肤创伤后1、7、14、21d观察创面愈合情况。结果提取的骨髓成体干细胞可以成功进行成骨、成脂诱导分化。骨髓移植后放射损伤动物全部存活;皮肤创伤后7d局部创面组织中可以观察到BrDU阳性细胞。皮肤创伤后14d,创面仅残留少量缺损组织,21d创面完全愈合。结论骨髓成体干细胞在皮肤创伤愈合过程中具有动员及募集作用,提高募集效率可能有助于改善创伤愈合能力。
【Abstract】 Objective To observe the mobilization and recruitment effect of bone marrow derived stem cells(BMSCs) in the repair of dermal wound for the advanced study on the mechanism of BMSCs in wound healing.Methods BMSCs were isolated from the bone marrow of male C57 mice,and purified by the differential adherence of different kinds of cells.The third generation of BMSCs were used for inducing osteogenesis and adipogenesis.In the meantime,the another portion of the third generation of BMSCs was labeled with 5-Bromo-2′-deoxyuridine(Brdu),then digested by trypsin and suspended in culture medium.The concentration of BMSCs was then adjusted to 108cells/ml.After 5Gy irradiation,6 female C57 mice were transplanted by way of tail vein with male BMSCs labeled with Brdu to establish the chimeric model of marrow transplantation.A dorsal skin defect was made in each of the marrow-chimeric mice 3 days after irradiation.A piece of wound tissue was harvested 7 days after operation in order to observe if the Brdu positive cells were recruited in the defect area.The wound surface was examined on days 1,7,14 and 21 after injury.Results Osteogenesis and adipogenesis were successfully induced from C57 mice BMSCs in vitro.The irradiated mice survived after BMSCs transplantation.The positive Brdu cells were observed in the wound tissue on day 7 post-operation.Only a small defect was found in the wound area 14 days after skin defects were made.The skin wound healed completely on day 21 post-operation.Conclusions BMSCs possess mobilization and "homing" effects in the wound area,and enhancement of these effects may be helpful for tissue repair.
【Key words】 adult stem cells; wounds and injuries; wound healing; recruitment;
- 【文献出处】 解放军医学杂志 ,Medical Journal of Chinese People’s Liberation Army , 编辑部邮箱 ,2009年09期
- 【分类号】R641
- 【下载频次】163