聚合物／液晶细胞模型的构建及其表面弹性对大鼠骨髓干细胞黏附的影响

Development of cell model of polymer/liquid crystal and effect of their elasticity on adhesion of rBM-MSCs

目的:构建聚合物/液晶细胞模型,探究其表面弹性对大鼠骨髓干细胞黏附情况的影响。方法:溶剂挥发致相分离方法构建聚合物/液晶细胞模型,通过偏光显微镜、扫描式电子显微镜及X射线衍射观察细胞模型的表面特征。全骨髓细胞贴壁培养法分离大鼠骨髓间充质干细胞(BMSCs),通过表面因子检测和诱导其成骨成脂分化进行鉴定。培养3代后分4组:纯PU空白对照组,10%膜组,30%膜组和50%膜组。接种细胞密度为5×107/L,待细胞贴壁24h后作细胞骨架染色处理,激光共聚焦显微镜观察细胞附着在材料上的情况并拍照。结果:溶剂挥发致相分离方法成功构建3种不同硬度的聚合物/液晶细胞模型;表面因子检测显示CD29、CD44、CD90呈阳性表达,而CD34和CD45呈阴性表达;经过茜素红S染色和油红O染色,成骨成脂分化形成的钙结节和脂滴都显而易见;细胞骨架染色拍摄结果显示,纯PU对照组、10%膜组和30%膜组上的细胞铺展面积较广,肌动蛋白微丝和细胞核清晰明显;而50%膜组的细胞铺展面积较小,细胞核虽较清晰,但是肌动蛋白微丝却模糊不清。结论:合适液晶含量的聚合物/液晶生物膜能支持大鼠骨髓干细胞的贴壁生长,而液晶含量过高则抑制细胞的贴壁生长。

AIM：To develop the cell model of polymer/liquid crystal and to study the effect of their elasticity on the adhesion of rat bone marrow mesenchymal stem cells （rBM-MSCs）.METHODS： Using the method of solvent e-vaporation induced phase separation, the cell model of polymer/liquid crystal was constructed.The surface morphology and phase separation structure were determined by polarized optical microscopy （ POM） , scanning electron microscopy （ SEM） and small angle X-ray scattering （ SAXS ） .rBM-MSCs were separated and expanded by adherent culture.The surface markers of rBM-MSCs were detected by flow cytometry.The cells were induced to osteogenic differentiation and adipogenic differentiation for 2 weeks.After 3 passages, the cells were divided into 4 groups, including total PU control group, 10%membrane group, 30%membrane group and 50%membrane group.The cells were then incubated with rhodamine phalloi-din for cytoskeleton staining and were observed under the confocal laser scanning microscope after cultured for 24 h.RE-SULTS：The cell model of polymer/liquid crystal was constructed successfully using the method of solvent evaporation in-duced phase separation.Flow cytometry results showed that the rBM-MSCs positively expressed CD29, CD44 and CD90, and negatively expressed CD34 and CD45.After stained with alizarin red S and oil red O, the calcium nodule and lipid droplets in rBM-MSCs were observed obviously.The cytoskeleton staining result indicated that the area in total PU control group, 10%membrane group and 30%membrane group were greater, and the actin microfilaments were also clearer than 〈br〉 that in 50%membrane group.CONCLUSION：The cell model with suitable content of liquid crystal made a contribution to the rBM-MSCs’ adhesion, but too much liquid crystal inhibits cell adhesion.