Jin Xuhong, Yang Liu, Zhang Shou, Dun Xiaojun, Wang Fuyou, Tan Hongbo
Affiliated Haikou Hospital, Xiangya School of Medicine, Central South University, Haikou 570203, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2012 Feb;29(1):125-8, 133.
The aim of this study was to label rabbit bone derived mesenchymal stem cells (BMSCs) with superparamagnetic iron oxide particles (SPIO) and to study the effects of magnetic labeling on the multi-differentiation of BMSCs. Rabbit BMSCs were isolated, purified, expanded, then coincubated with SPIO(25 microg/ml) complexed to protamine sulfate (Pro) transfection agents overnight. Prussian blue staining and transmission electron microscopy were performed to show intracellular iron. Cell differentiation was evaluated. Both labeled and unlabeled BMSCs were subjected to osteogenic, adipogenic and chondrogenic differentiation to assess their differentiation capacity for 21 d. Osteogenic cells were stained with alizarin red to reveal calcium deposition, adipogenic cells were stained with oil redO' respectively. Chondrogenic cells stained with Safranin-O, glycosamino glycans, and type II collagen production was assessed by standard immunohistochemistry. Cell with immunohistochemistry staining were detected by polarized light microscopy and analysed by Image-Pro Plus software. The results showed that intracytoplasmic nanoparticles were stained with Prussian blue and observed by transmission electron microscopy clearly except the unlabeled control. As compared with the nonlabeled cells, it showed no statistically significant difference on the differentiation of the labeled BMSCs. And the differentiation of the labeled cells were unaffected by the endosomal incorporation of SPIO. In summary, BMSCs can be labeled with SPIO without significant change in cell multi-differentiation capacity.
本研究旨在用超顺磁性氧化铁颗粒（SPIO）标记兔骨髓间充质干细胞（BMSCs），并研究磁性标记对BMSCs多向分化的影响。分离、纯化、扩增兔BMSCs，然后与硫酸鱼精蛋白（Pro）转染剂复合的SPIO（25μg/ml）过夜共孵育。进行普鲁士蓝染色和透射电子显微镜检查以显示细胞内铁。评估细胞分化。对标记和未标记的BMSCs进行成骨、成脂和软骨分化，以评估其21天的分化能力。成骨细胞用茜素红染色以显示钙沉积，成脂细胞分别用油红O染色。软骨细胞用番红O染色，通过标准免疫组织化学评估糖胺聚糖和II型胶原蛋白的产生。通过偏振光显微镜检测免疫组织化学染色的细胞，并使用Image-Pro Plus软件进行分析。结果显示，除未标记的对照外，细胞质内纳米颗粒经普鲁士蓝染色，透射电子显微镜可清晰观察到。与未标记细胞相比，标记的BMSCs在分化方面无统计学显著差异。并且标记细胞的分化不受SPIO内吞作用的影响。总之，BMSCs可用SPIO标记，而细胞多向分化能力无明显变化。
