摘要
通过DiI荧光标记后的骨髓基质干细胞(BMSC)与β-磷酸三钙(β-TCP)支架复合后的镜下观察,找到一种大块支架材料上活细胞观察的新方法。为组织工程骨的研究提供新的可行性方法。取犬的第3代BMSC消化后分两组,实验组和对照组,实验组以DiI标记,另一组正常细胞不标记,测定生长曲线。另以相同数量两组细胞同β-TCP材料复合后培养,测定两组细胞贴附率,在第3、7、14天时,测其在β-TCP材料上的增值情况,并在荧光显微镜下连续观察标记后的BMSC的生长状况。结果表明,实验组和对照组的生长曲线基本吻合,两组之间各时间点的细胞计数都没有统计学意义(P>0.05)。两组细胞与β-TCP材料复合三维共培养后,第3、7、14天细胞增值率未见显著性差异(P>0.05)。荧光显微镜观察染色后的BMSC细胞可见细胞胞膜发出红色荧光呈环状,胞核未着色,显现良好。DiI荧光标记后的BMSC生长增殖,可在荧光显微镜下活体连续观察,在大段不透明支架材料上的细胞观察能作为一种较好的新方法来研究种子细胞与支架材料的相互作用,可以成为组织工程骨研究中的新观察方法推广应用。
To find out a novel method of investigating live cells on great scaffold material, through observing the combination of BMSC labeled by DiI and β-TCP scaffold,and to offer a new feasible method to study the tissue engineering bone, the third passage BMSC of dogs digested were divided into two groups： the experiment group was labeled by DiI, and the normal group not. They were planted into cell culture plates with 12 holes at the same level, each group with 6 holes per plate. From then on, one plate each day is took to digest, counted cells for 7 days continuously and drew the growth curve. Otherwise, two group ceils of the same number combined with the β-TCP scaffold are made, then cultured and evaluated their adhesion rate. Digesting and counting each group cells on the day of 3,7, 14, the increasing state with the β-TCP scaffold is observed, and investigated the growth state of the DiI labeled cells by fluorescent microscope. The results showed that the growth curve of each group is not significant on the whole, and so are the cell numbers on different time（P〉0.05）. The increasing rates of the two groups on the β-TCP scaffold are not significant on the day of 3,7,14 （P〉0.05）. BMSC labeled by DiI can be observed clearly by fluorescent microscope. Compared with the normal cells, the proliferation and appearance of BMSC labeled by DiI are not significant. The BMSC labeled by DiI can be well compatible with β-TCP scaffold and be observed continuously in living state. The novel method of observation the cells on the opaque scaffold is feasible to investigate the interaction between the cells and the opaque great scaffold. The method can be utilized extensively in the tissue engineering bone.
出处
《科学技术与工程》
2007年第2期164-168,共5页
Science Technology and Engineering
基金
国家自然科学基金重点项目(50235020)资助
作者简介
王超锋，第四军医大学西京医院骨科研究所研究生．研究方向：骨肿瘤，组织工程骨。
通讯作者简介：王臻，第四军医大学西京医院骨科研究所主任医师。
