Liu Weidong, Lu Guohua, Wang Bing, Ma Zemin, Li Yawei
Department of Spine Surgery, Second Xiangya Hospital, Central South University, Changsha, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2012 May;37(5):441-6. doi: 10.3969/j.issn.1672-7347.2012.05.002.
To provide further foundation for the therapy for spine cord injury by transferred mesenchymal stem cells (MSC) expressing the brain-derived neurotrophic factor (BDNF) gene.
Bone mesenchymal stem cells were isolated from rats and directly cultivated and expanded in vitro. Bone mesenchymal stem cells were divided into 3 groups: a pEGFP-N1- BDNF-transfected group (BDNF group), a pEGFP-N1 transfected group (vector group) and a non-transfected group. Western blot was applied to detect the expression of transgenic bone mesenchymal stem cells. The differentiation of bone mesenchymal stem cells was identified by immunofluorescence. The rates of induction of neuron-like cells among the three cell groups were compared.
The cell surface markers of bone mesenchymal stem cells included expression of CD90 (+), CD44 (+), CD34 (-), CD45 (-). The expression of BDNF in the transfected bone mesenchymal stem cells was demonstrated by Western blot. The positive ratio of neuron-like differentiation in the BDNF group was higher than that of both the vector group and the non-transfected group (P<0.05).
BDNF gene plays an important role in promoting bone marrow mesenchymal stem cells to differentiate into neuron-like cells.
为通过转染表达脑源性神经营养因子（BDNF）基因的间充质干细胞（MSC）治疗脊髓损伤提供进一步的理论基础。
从大鼠中分离出骨髓间充质干细胞并在体外直接培养和扩增。将骨髓间充质干细胞分为3组：pEGFP-N1-BDNF转染组（BDNF组）、pEGFP-N1转染组（载体组）和未转染组。采用蛋白质印迹法检测转基因骨髓间充质干细胞的表达。通过免疫荧光鉴定骨髓间充质干细胞的分化情况。比较3组细胞中诱导神经元样细胞的比率。
骨髓间充质干细胞的细胞表面标志物包括CD90（+）、CD44（+）、CD34（-）、CD45（-）的表达。蛋白质印迹法证实了转染的骨髓间充质干细胞中BDNF的表达。BDNF组神经元样分化的阳性率高于载体组和未转染组（P<0.05）。
BDNF基因在促进骨髓间充质干细胞向神经元样细胞分化中起重要作用。
