黄芪诱导大鼠骨髓间充质干细胞分化为神经样细胞的研究

目的研究大鼠骨髓间充质干细胞(MSCs)体外分离和培养方法,并用黄芪体外诱导骨髓间充质干细胞分化为神经细胞样细胞。方法利用梯度离心从大鼠骨髓中分离单核细胞进行培养,去除不贴壁的细胞,分离纯化,对其生长特性进行分析。采用含黄芪的无血清L-DMEM诱导分化为神经细胞样细胞,观察细胞形态变化,用免疫组织化学方法检测分化细胞中巢蛋白(nestin)、神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)的表达。结果用含10%胎牛血清(FCS)的培养基培养,原代细胞贴壁生长,细胞形态均一,为纺锤形,有克隆团形成。传代培养时,形态变为成纤维细胞样,有较强的增殖能力。经黄芪诱导后,骨髓间充质干细胞形态发生改变,nestin、NSE和GFAP阳性,分化为神经元或胶质细胞样细胞。结论骨髓间充质干细胞可以体外分离、培养,在一定条件下向神经样细胞分化。

Study on Inducing Effect of Astragalus mongholicus in Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells into Neurocyte-Like Cells

Objective To investigate differentiation of rat bone marrow-derived mesenchymal stem cells(MSCs)into neurocyte-like cells inducing by Astragalus mongholicus.Methods we isolated the nucleated cells from rat bone marrow through gradient centrifugation,then cultured them and removed the unadherent cells,then got pure MSCs to study their growth character.Astragalus mongholicus contained in non-serum-L-DMEM was used to induce differentiation of MSCs into neurcyte-like cells,and then observed morphologic changes of differentiated cells,identified them detecting the nestin,neuron specific enolase(NSE) and glial fibrillary acidic protein(GFAP) by immuncytochemistry.Results The primary cells grew well in adherent way when cultured in L-DMEM containing 10% fetal calf serum(FCS),and were fusiform shape,then could form cell clones.Subculturing cells were fibroblast-like,and had higher reproductive activity than the primary cells.Morphology of MSCs change when they cultured with Astragalus mongholicus.Nestin,NSE and GFAP were positive,MSCs became neuron-like and gliocyte-like cells.Conclusion MSCs can be isolated and cultured in vitro.If the culture is appropriate,it can diffenentiate into neurocyte-like cells.