CAJ | 학술논문

目的：使用补肾益脑胶囊将大鼠骨髓间充质干细胞(rMSC)体外诱导为神经细胞。方法：体外扩增培养大鼠股骨骨髓细胞。MSC通过补肾益脑胶囊内药粉溶解液含10%胎牛血清的L-DMEM预诱导24h后，用无血清L-DMEM内加入补肾益脑胶囊内药粉溶解液以正式诱导 MSC 分化成神经细胞。通过倒置显微镜对比观察细胞前后的形态变化；神经细胞特征性结构则以尼氏染色法来观察；以免疫细胞化学鉴定有神经干细胞标志物巢蛋白(nestin)、胶质纤维酸性蛋白(GFAP)和神经元烯醇化酶(NSE)的表达。结果：通过贴壁法成功将大鼠骨髓间质干细胞分离并在体外大量扩增，补肾益脑胶囊诱导3小时后，MSC形态转变为典型的神经样细胞，尼氏染色法显示诱导后的细胞胞体中有尼氏体，免疫细胞化学染色鉴定显示其NSE、nestin表达皆为阳性，GFAP表达阴性。结论：补肾益脑胶囊诱导下大鼠骨髓间质干细胞可分化为多种形态的神经样细胞。
목적：사용보신익뇌효낭장대서골수간충질간세포(rMSC)체외유도위신경세포。방법：체외확증배양대서고골골수세포。MSC통과보신익뇌효낭내약분용해액함10%태우혈청적L-DMEM예유도24h후，용무혈청L-DMEM내가입보신익뇌효낭내약분용해액이정식유도 MSC 분화성신경세포。통과도치현미경대비관찰세포전후적형태변화；신경세포특정성결구칙이니씨염색법래관찰；이면역세포화학감정유신경간세포표지물소단백(nestin)、효질섬유산성단백(GFAP)화신경원희순화매(NSE)적표체。결과：통과첩벽법성공장대서골수간질간세포분리병재체외대량확증，보신익뇌효낭유도3소시후，MSC형태전변위전형적신경양세포，니씨염색법현시유도후적세포포체중유니씨체，면역세포화학염색감정현시기NSE、nestin표체개위양성，GFAP표체음성。결론：보신익뇌효낭유도하대서골수간질간세포가분화위다충형태적신경양세포。
Objective:To investigate the differentiation from rat mesenchymal stem cells (rMSCs)to neuron-like cells with Bushen Yinao Capsules. Methods: Mesenchymal stem cells were obtained from thighbones marrow of rats and expended in culture medium. Bushen Yinao Capsules was used to study its differentiative effect on MSCs. MSCs were pretreated with preinduction medium consisting of L-DMEM/10%FCS/Bushen Yinao Capsules for 24 hours,then they were induced with induction media consisting of L-DMEM/Bushen Yinao Capsules.The morphologic changes were observed by light microscope,the specific characteristic structures of neurons cells were observed using Nissl’s staining method,Neuron-specific enolase(NSE),nestin,glial fiber acidic protein(GFAP)were detected by immunocytochemistry. Results:Rat MSCs were expanded as undifferentiated cells . MSCs were induced with Bushen Yinao Capsules, which could generate neuron-like cells within 3 hours. Trachychromatic Nissl body was found in cytolymph.The expression of NSE and nestin in the neuron-like cells was positive but GFAP was negative. Conclusion:MSCs can be induced to differentiate into neuron like-cells with the effect of Bushen Yinao Capsules.