移植骨髓间充质干细胞后的脑脊液促进神经干细胞的分化

目的观察骨髓间充质干细胞（BMSC）移植于损伤脊髓后，所提取的脑脊液（BMSCs-CSF）对体外培养的神经干细胞向神经元细胞分化的影响，探讨BMSCs促进神经干细胞分化的机制。方法培养BMSCs及神经干细胞；制作大鼠脊髓损伤模型；将体外培养的BMSCs注入脊髓损伤区，于术后3d、5W分别提取脑脊液，并注入神经干细胞培养液中，观察神经干细胞向神经元细胞的分化。试验分为五组，A：无脊髓损伤组脑脊液；B1：脊髓损伤3天后收集的脑脊液；B2：脊髓损伤组5周后收集的脑脊液；C1：脊髓损伤并行BMSCs移植，在3天后收集的脑脊液；Q2：脊髓损伤并进行BMSCs移植，在5周后收集的脑脊液。结果加入脑脊液培养72小时后在倒置显微镜下观察细胞形态结果显示A、B1、B2各组神经干细胞球无贴壁，悬浮于培养基中，神经干细胞无细胞突形成；C1组显示神经干细胞球贴于培养板底部，并有细胞突生长。C2组显示仅有少量的神经干细胞球贴壁，细胞突短小少见。结论BMSCs移植于损伤的脊髓后所提取的脑脊液（BMSCs-CSF）具有促进神经干细胞向神经元细胞分化的作用。

Research on the effect of BMSCs-CSF on the differentiation of neurosphere cells

Obkective To study the effect of BMSCs-CSF（CSF from injured spinal cord treated by transplantation d BMSCs） on the differentiation of neurosphere cells.Methods BMSCs and Neurosphere cells were cultured; 50 SD rats were randomly assigned into 5 groups.In groupA,the spinal cord were exposed and keep the cord intact,and CSF was collected from the cisterna magna. In groupBland B2,SCI models were made,and CSF was collected 3d and 5w postopertation respectively.In groupC1 and C2,SCI modela were treated with BMSCa transplantation,and CSF was collected 3 d and 5w postoperation respectively. Neurosphere cells were treated with CSF of each group respectively. The change of neurospheres including adhesion to the culture dish and cellular process extension were observed using a phase contrast microscope 3 days after CSF added. Immunohistochemical study was used to analysis the immunoreactivity to NF. Results In groupA, B1 and B2, the neurosphere cells did not adhere to the dish bottom and no extended cellular processes were observed. In GroupC1, the neurosphere cells adhered to the dish bottom and long cellular processes extended toward the periphery. In GroupC2, the nettrosphere ceils adhered to the dish bottom and no long cellular processes extended. NF staining was positive in groupC1. Conclusion BMSCs-CSF can improve the differentiation of neurosphere cells toward neurons.