体外诱导骨髓间充质干细胞向神经元样细胞分化：两种方法的比较

摘要

背景：目前骨髓间充质干细胞诱导分化为神经细胞的方法较多，而采用不同诱导方法时间充质干细胞在体外分化成神经细胞的比例是不一样的。适宜的诱导条件是实现间充质干细胞定向诱导分化的必要条件。目的：比较两种常见的化学诱导法与共培养法诱导大鼠骨髓间充质干细胞向神经细胞分化的差异，以寻找一种诱导效果高、实用的骨髓间充质干细胞体外诱导方法。方法：采用密度梯度法分离培养大鼠骨髓间充质干细胞，分别用化学诱导法和共培养法诱导分化比较两种方法所获得的神经细胞数目、细胞形态和特异性抗体阳性率。结果与结论：培养7 d 后两组均可见大量贴壁细胞形成突起，呈放射状生长，神经元特异性烯醇化酶染色均为阳性。共培养法第5天可见典型神经细胞结构，突起数量较多，神经元特异性烯醇化酶染色阳性率(50.82±2.46)%，化学诱导法培养第7天可见神经样细胞形成，并有突起，神经元特异性烯醇化酶染色阳性率(43.56±1.74)%。说明骨髓间充质干细胞经共培养法诱导分化后的神经样突起数量多并较早互相形成连接，且共培养法神经元特异性烯醇化酶染色阳性率高于化学诱导法。%BACKGROUND: A variety of methods have been used to induce bone marrow mesenchymal stem cells to differentiate into nerve cells. However, the percent of differentiated nerve cells is different using varied induction methods for different time periods. Appropriate induction condition is essential for induced differentiation of mesenchymal stem cells. n OBJECTIVE: To find a suitable and practical method of inducing bone marrow mesenchymal stem cells into nerve cells in vitro by comparing chemical induction and co-culture methods. n METHODS: Rat bone marrow mesenchymal stem cells were isolated by density gradient method and cultured by chemical induction and co-culture methods, respectively. Cell number, cell morphology, and specific cell surface markers were compared. n RESULTS AND CONCLUSION: A large number of adherent cells with processes and radial growth, positive for neuron specific enolase staining were observed in both groups. Cells with typical nerve cell structure and a large number of processes were observed at 5 days in co-culture group, and the positive rate for neuron specific enolase staining was (50.82±2.46)%. Neural-like cells with processes were found at 7 days in chemical induction group, and the positive rate for neuron specific enolase staining was (43.56±1.74)%. Results showed that bone marrow mesenchymal stem cells were inducted into neural-like cells with more processes, which formed connection earlier cultured in co-culture compared with chemical induction. Moreover, the positive rate for neuron specific enolase staining was higher in co-culture group compared with chemical induction group.