Li Wen-yu, Jin Ri-long, Hu Xing-yue
Psychology Department, Zhejiang University, Hangzhou, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2012 Nov;41(6):659-64. doi: 10.3785/j.issn.1008-9292.2012.06.009.
To investigate the migration of fluorescent dye PKH26-labeled BM-MSC in the Alzheimer's model rats.
Normal human bone marrow extracted for isolation of BM-MSC was cultured in vitro. The 5th passaged BM-MSC was labeled with PKH26, and observed under a fluorescence microscope for PKH26 labeling efficiency, and using flow cytometry BM-MSC surface markers was checked. The PKH26 labeled BM-MSC injected into the tail vein of the normal control group and AD animal model group, 14 days after finding the PKH26-labeled BM-MSC cells in the rat hippocampus using fluorescence microscopy. Using the Morris water maze experiment comparison of AD model and BM-MSC transplantation group of spatial learning and memory ability.
TFlow cytometry showed BM-MSC surface markers CD73 and CD105 were positive. In vitro, PKH26-labeled rate of BM-MSC was 100 %. The Morris water maze experiment comparison of BM-MSC transplantation group and AD group of animals, BM-MSC transplantation group at 13, 14 days of spatial learning and memory ability than AD animal group had significantly improved. 14 days after BM-MSCs in rat hippocampus could be found which were PKH26-positive, consistent with DAPI staining. PKH26-positive cells in animal models of AD were significantly more than those in the normal control group.
BM-MSC in AD rats not only migrates through the blood-brain barrier, but also mainly survives in the hippocampus of AD rats, and it can improve AD rat model of learning disabilities.
研究荧光染料PKH26标记的骨髓间充质干细胞(BM-MSC)在阿尔茨海默病模型大鼠中的迁移情况。
提取正常人骨髓用于分离BM-MSC并进行体外培养。第5代BM-MSC用PKH26标记,在荧光显微镜下观察PKH26标记效率,并用流式细胞术检测BM-MSC表面标志物。将PKH26标记的BM-MSC注入正常对照组和AD动物模型组的尾静脉,14天后用荧光显微镜在大鼠海马中寻找PKH26标记的BM-MSC细胞。采用Morris水迷宫实验比较AD模型组和BM-MSC移植组的空间学习和记忆能力。
流式细胞术显示BM-MSC表面标志物CD73和CD105呈阳性。体外实验中,BM-MSC的PKH26标记率为100%。Morris水迷宫实验比较BM-MSC移植组和AD组动物,BM-MSC移植组在第13、14天的空间学习和记忆能力比AD动物组有显著提高。BM-MSC注入大鼠体内14天后,在海马中可发现PKH26阳性细胞,与DAPI染色结果一致。AD动物模型中PKH26阳性细胞明显多于正常对照组。
AD大鼠中的BM-MSC不仅能穿越血脑屏障,还主要存活于AD大鼠的海马中,且能改善AD大鼠模型的学习障碍。