【摘要】 目的探讨丹参注射液对人脐血单个核细胞来源的间充质干细胞(MSCs)向神经细胞分化的诱导作用。方法利用FACScan流式细胞仪检测MSCs表面抗原CD29、CD44、CD59、CD33,用丹参注射液诱导人脐血原代细胞和脐血来源的间充质干细胞向神经细胞方向分化,并与神经生长因子(NGF)和神经常苷脂(GM1)的诱导作用比较,用免疫细胞化学方法对分化和未分化细胞进行鉴定。结果人脐血原代细胞中MSCs的表耐标记CD29、CD44、CD59阳性率分别为10.7%、37.27%和66.67%,而造血细胞的表面标记CD33阳性率仅为0.33%。人脐血传代细胞(第5代)MSCs的表面标记CD29、CD44、CD59的阳性率分别为40.2%、70.5%和95.4%。原代培养的贴壁细胞形态呈大小不等圆形和条形,用丹参诱导可表达神经细胞的标记。传代培养的间充质干细胞,可维持在未分化状态稳定增殖。用丹参可诱导这种细胞向神经样细胞分化,表达神经干细胞标记nestin,神经元的标记β-Ⅲ类神经微管(β-TubulinⅢ)、神经微丝(NF)和神经胶质细胞的标记胶质纤维酸性蛋白(GFAP)。与NGF和GMI的诱导作用比较,细胞形态类似,表达相同的神经细胞标记,丹参的诱导速度较快,诱导后细胞表达神经元标记的比例较高。结论人脐血是MSCs的来源之一,丹参可诱导人脐血干细胞分化为神经样细胞,丹参可作为神经诱导剂,人脐血干细胞可作为神经干细胞的来源。更多还原

【Abstract】 Objective To investigate the differentiation - inducing effects of salvia miltiorrhiza on umbilical cord blood mesenchy-mal stem cells. Methods The immunophenotypes of human umbilical cord blood-derived mesenchymal stem cells(MSCs) were determined by flow - cytometry. Salvia miltiorrhiza was adapted to induce umbilical cord blood primary cultured cells and MSCs to differen tiate. The differentiated and undifferentiated cells were identified by immunocytochemistry. Results In primary culture, a few cells proliferated and formed cell clusters. These cells were big or small round shapes and strip shapes. The positive rates for MSCs phenotypes CD29 , CD44 , CD59 of primary culture cells were 10.7% ,37.27% and 66.67%, respectively. Salvia miltiorrhiza could induce the umbi lical cord blood primary cultured cells to express neural phenotypes. After passage, the MSCs could be maintained in vitro for extended periods with stable population doubling. The positive rates for CD29, CD44, CD59 of human umbilical cord blood-derived MSCs were 40.2% ,70.5% and 95.4% , respectively. Salvia miltiorrhiza could induce the MSCs to express three kinds of proteins: nestin (the marker of neuronal precursor stem cells) ,β- TubulinIII and neurofilament (NF) (the marker of neuronal phenotypes) and glial fibrillary acidic protein(GFAP, the marker of astrocyte). Compared with nerve growth factor(NGF) and ganglioside(GM1) ,the induction effect of salvia miltiorrhiza demonstrated the higher rate of expressing neuron phenotypes at a faster speed.Conclusions The human MSCs could be isolated from human umbilical cord blood. Salvia miltiorrhiza can induce the MSCs to differentiate into nerve- like cells, therefore, might act as a potential drug for neural induction.更多还原