小鼠非黏附骨髓间充质干细胞分化为神经元样细胞的实验研究  

Non-adherent bone marrow-derived mesenchymal stem cells differentiate into neuron-like cells in mice

作　　者：王燕[1] 陈燕玲[1] 贲晓明[1] 周晓玉[1] 苗登顺[2] 

机构地区：[1]南京医科大学附属南京儿童医院新生儿科,210008 [2]南京医科大学骨与干细胞研究中心

出　　处：《中华围产医学杂志》2010年第2期123-127,共5页Chinese Journal of Perinatal Medicine

基　　金：国家自然科学基金（30471836）

摘　　要：目的探讨小鼠非黏附骨髓间充质干细胞（non-adherent bone marrow-derived mesenchymal stem cell，NA—BM—MSC）在体外、体内尤其是缺血损伤的脑内向神经元样细胞分化的能力。方法传代纯化后的NA—BM-MSC经表皮生长因子和碱性成纤维细胞生长因子诱导后，通过甲苯胺蓝染色和免疫细胞化学染色检测神经元特异性核蛋白和神经中间丝蛋白的表达情况；将来自β-半乳糖苷酶转基因小鼠的NA—BM-MSC在立体定位仪下移植到小鼠缺血损伤的脑内，8周后通过LacZ组织化学染色和免疫组织化学染色检测供体细胞在缺血脑内微环境中存活以及神经元特异性核蛋白的表达情况。结果NA—BM-Msc经过诱导能够表达神经元特异性核蛋白和神经中间丝蛋白并有尼氏体形成；移植到损伤脑内后，LacZ组织化学染色发现在缺血侧供体细胞能够存活，免疫组织化学单、双染显示在缺血坏死区及周围都检测到了β-半乳糖苷酶阳性的供体细胞，其中的部分阳性细胞还同时表达神经元特异性核蛋白。结论NA-BM-MSC在体外能够分化成神经元样细胞，在小鼠缺血损伤的脑组织内也能够存活、迁移，部分细胞还能分化为神经元样细胞。Objective To determine whether non-adherent bone marrow-derived mesenchymal stem cells （NA-BM-MSC） can differentiate into neuron-like cells in vitro and in vivo, especially in ischemic brain of mice. Methods NA-BM-MSC of mouse were induced with conditioned medium containing epidermal growth factor and basic-fibroblast growth factor and demonstrated by immunocytochemistry for the expression of neuron specific nuclear protein and neurofilament-200. NA-BM-MSC from β-galactosidase transgenie mice constitutively expressing β-galactosidase were transplanted into the mice model of middle cerebral artery occlusion. LaeZ staining and immunohistochemistry staining were performed to detect the survival, distribution and the differentiation of the donor cells in the ischemic brain. Results NA-BM-MSC can be induced into neuron specific nuclear protein and neurofilament-200 positive cells in vitro. LacZ staining showed that NA-BM-MSC can survive in ischemic brain and express β-galactosidase. Also,numbers of the neuron specific nuclear protein positive cells in β-galactosidase-positive cells were detected in the ischemic brain with double immunohistochemistry staining. Conclusions NA-BM-BMC can be induced to differentiate into neuron-like cells in vitro, and also can differentiate into neuron-llke ceils in ischemic brain tissue of the mice.

关 键 词：间质干细胞 细胞分化 神经元 脑缺血 问质干细胞移植 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]