人雪旺细胞促进人脐带间充质干细胞向神经方向分化的研究  

Neural differentiation of human umbilical cord mesenchymal stem cells promoted by uman schwann cells

作　　者：陈有[1] 冯世庆[1] 吴秋丽[1] 周先虎[1] 刘举[1] 宁广智[1] 冯洪永[1] 周恒星[1] 

机构地区：[1]天津医科大学总医院骨科,300052

出　　处：《中华实验外科杂志》2012年第5期930-932,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30872603）;天津市卫生局科技重点资金资助项目（07KG2）;天津市卫生局科研基金资助项目（09KZ104）

摘　　要：目的观察人雪旺细胞（hSCs）对人脐带间充质干细胞（hUC—MSCs）在脊髓损伤（SCI）体内存活、分化的影响，通过分子生物学技术检测胶质纤维酸性蛋白（GFAP）、髓磷脂碱性蛋白（MBP）与高分子量神经丝蛋白（NF—H）的表达，描述其变化的时间特征。方法分离、培养及纯化hUC。MSCs和hSCs。取8周龄雌性Wistar大鼠60只，以ImpactorModel-Ⅱ型打击器制作脊髓胸10（TIO）损伤模型。大鼠随机均分为4组：DMEM对照组（A）、hSCs移植组（B）、hUC—MSCs移植组（C）、hUC—MSCs与hSCs联合移植组（D），各组分别于移植后1、2、3、4周取材。结果D组可见MBP与核因子（NF）-H染色阳性细胞，其他各组为阴性。A组3种神经标志物在蛋白与mRNA水平的表达量随时间延长均逐渐增高，且GFAP在移植后2周明显增高，MBP与NF-H在移植后3周明显增高[2周与1周GFAP条带灰度比值为1．34，而聚合酶链反应（PCR）所测比值为2．86；3周与1周MBP和NF．H条带灰度比值分别为3．43、4．12，而PCR所测比值分别为6．47、6．78]。与对照组相应时间点比较。移植组MBP、NF．H的表达量较高而GFAP的表达量较低，其中D组的MBP、NF—H的表达量最高（D组与A组MBP3周条带灰度比值为2．11，而NF—H3周条带比值为2．11），差异有统计学意义（P〈0．05），但D组GFAP各时间点的表达量差异（各周所测条带灰度比值依次为1．00：1．12：1．13：1．14）无统计学意义（P〉O．05）。结论hSCs可促进hUC—MSCs向神经元与少突胶质细胞方向分化并增强其对胶质瘢痕的抑制作用。Objective To discuss the＇influence of human schwann cells （hSCs） on the survival and differentiation of human umbilical cord mesenchymal stem cells （hUC-MSCs） in Wistar rats with spinal cord injury （SCI） ; To detect the expression of glial fibrillary acidic protein （GFAP）, myelin basic protein （MBP） and high molecular weight neurofilament （NF-H）, and describe their changes of the time-depend- ent characteristic. Methods Firstly, the hUC-MSCs and the hSCs were isolated and purified in vitro. Sixty 8-week-old female Wistar rats were subjected to T10 SCI by Impactor Model- ]] type device and divided in- to four groups randomly and evenly： DMEM control group （A）, hSCs transplantation group （B）, hUC-MSCs transplantation group （ C）, co-transplantation group of hSCs and hUC-MSCs （D）. The injured spinal cord tissues were taken out at 1st, 2nd, 3rd, and 4th week after cell transplantation. Results The MBP and NF-H positive ceUs could be seen in group D, but not in other groups. The three neural markers in group A were gradually increased with time, and the expression of GFAP was significantly increased at the 2nd week following transplantation, while the rest two markers were significantly increased at the 3rd week （The GFAP band gray-value ratio at 2nd week to that at 1st week was 1.34： 1.00, while PCR ratio was 2. 86： 1.00; The MBP band gray-value ratio at 3rd week to that at 1st week was 3.43： 1.00, while PCR ratio was 6.47： 1.00; For NF-H, 4. 12：1.00 and 6. 78：1.00 respectively）. As compared with the group A at the corresponding time point, the expression of both MBP and NF-H was increased in groups B, C and D, while the expression of GFAP was reduced, more precisely, the expression of MBP and NF-H was the highest in group D （The MBP and NF-H band gray-value ratio at 3rd week to that at 1st week in both group D and group A was 2. 11 ： 1.00, P 〈 0.05 ）. However, there was no significant change in the GFAP expression at each time point in group D （The

关 键 词：脊髓损伤 细胞移植 神经分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]