Jing Meng, Liu Xin-quan, Liang Peng, Li Chang-yu, Zhang Xiang-tong, Wang Dan, Luan Yu, Liu En-zhong
Department of Neurosurgery, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, China.
Zhonghua Yi Xue Za Zhi. 2004 Aug 17;84(16):1386-9.
To evaluate the feasibility of monitoring the neural stem cells implanted into the brain by the technique of labeling with superparamagnetic iron oxide (SPIO).
Neural stem cells were isolated from the cerebral cortex of newborn Wister rats and cultured. SPIO particles and poly-L-lysine were added into the medium to be co-cultured foe one hour. After the formation of neurospheres, Prussian blue staining was conducted and transmission electron microscopy was used to identify the iron particles in these neural stem cells. Sixteen adult female Wistar rats underwent transplantation of labeled neural stem cells into the right side of brain and non-labeled cells were transplanted into the contralateral part as controls. 1, 2, 4, 6, and 7 weeks after the transplantation, MRI examination with the scanning sequences of SE T2WI, FSE T2WI, and GRE T2 * respectively was conducted on the brains of the rats. Four rats at each time point were killed and their brains were taken out to undergo HE staining and Prussian blue staining to track the presence of labeled-cells.
After the addition of SPIO the neurospheres continued to proliferate and differentiate normally. Electron microscopy showed vacuolar structures of different sizes under the cytoplasma membrane within and outside which there were high-density iron particles. Prussian blue staining showed numerous blue stained particles in the cytoplasm of the labeled cells. Remarkable low signal change was seen in the right brain transplanted with labeled cells, especially in the condition of scanning sequence of GRET2. Such change could be seen up to 7 weeks after the transplantation. No signal change was found in the left brain.
SPIO labeling technique is useful in monitoring the outcome of transplanted neural stem cells.
评估采用超顺磁性氧化铁(SPIO)标记技术监测植入脑内的神经干细胞的可行性。
从新生Wistar大鼠的大脑皮质分离神经干细胞并进行培养。将SPIO颗粒和聚-L-赖氨酸加入培养基中共培养1小时。形成神经球后,进行普鲁士蓝染色,并使用透射电子显微镜鉴定这些神经干细胞中的铁颗粒。16只成年雌性Wistar大鼠接受将标记的神经干细胞移植到右侧脑内,将未标记的细胞移植到对侧作为对照。移植后1、2、4、6和7周,分别对大鼠脑进行SE T2WI、FSE T2WI和GRE T2*扫描序列的MRI检查。每个时间点处死4只大鼠,取出其脑进行HE染色和普鲁士蓝染色以追踪标记细胞的存在情况。
加入SPIO后神经球继续正常增殖和分化。电子显微镜显示细胞膜下有不同大小的空泡结构,其内外有高密度铁颗粒。普鲁士蓝染色显示标记细胞的细胞质中有大量蓝色染色颗粒。在移植标记细胞的右侧脑内可见明显的低信号变化,尤其是在GRE T2扫描序列的情况下。这种变化在移植后7周仍可见。左侧脑未发现信号变化。
SPIO标记技术有助于监测移植神经干细胞的结果。