Porous chitosan microspheres with diameters ranging from 180μm to 280 μm were successfully prepared, using an anti-phase suspension method combined with temperature controlled freeze-extraction. The mean pore diamet...Porous chitosan microspheres with diameters ranging from 180μm to 280 μm were successfully prepared, using an anti-phase suspension method combined with temperature controlled freeze-extraction. The mean pore diameter could be regulated from 5 μm to 60μm by varying the freezing temperature through the cooling rate. Results with in vitro chondrocyte cultures showed that cells could attach, proliferate and spread on these porous microspheres as well as inside the microcarriers. The materials and cell cocultures were characterized using both optical and scanning electron microscopy. These results show that the porous chitosan microspheres are promising candidates for tissue culture for use as an injectable tissue engineering scaffold.展开更多
[目的]探索在旋转生物反应器内,应用微载体技术快速扩增并向软骨分化人脂肪干细胞。[方法]将人脂肪干细胞结合Cytodex3微载体在旋转的生物反应器(RCSS)内进行动态培养,应用倒置显微镜和扫描电镜对微载体表面的脂肪干细胞进行动态观察,...[目的]探索在旋转生物反应器内,应用微载体技术快速扩增并向软骨分化人脂肪干细胞。[方法]将人脂肪干细胞结合Cytodex3微载体在旋转的生物反应器(RCSS)内进行动态培养,应用倒置显微镜和扫描电镜对微载体表面的脂肪干细胞进行动态观察,并对收获的脂肪干细胞进行Safran in-O、tolu id ine b lue染色等组织化学染色及Ⅱ型胶原的免疫化学染色分析。[结果]脂肪干细胞于24 h内贴附于Cytodex3微载体表面,细胞形态为短梭形,随时间的延长,贴附于微载体的细胞逐渐增多,到培养后期,细胞密度可达最初接种的19倍左右,在微载体上收获的细胞进行番红花O、阿利新蓝染色呈阳性,Ⅱ型胶原染色阳性,均强于对照组。[结论]利用微载体细胞培养技术可简便快速地在体外扩增脂肪干细胞,并成功实现向软骨细胞分化。展开更多
基金Supported by the National Key Basic Research and Development (973) Program of China (No. 2005CB623905) and the Tsinghua-Yue-Yuen Medical Science Fund
文摘Porous chitosan microspheres with diameters ranging from 180μm to 280 μm were successfully prepared, using an anti-phase suspension method combined with temperature controlled freeze-extraction. The mean pore diameter could be regulated from 5 μm to 60μm by varying the freezing temperature through the cooling rate. Results with in vitro chondrocyte cultures showed that cells could attach, proliferate and spread on these porous microspheres as well as inside the microcarriers. The materials and cell cocultures were characterized using both optical and scanning electron microscopy. These results show that the porous chitosan microspheres are promising candidates for tissue culture for use as an injectable tissue engineering scaffold.
文摘[目的]探索在旋转生物反应器内,应用微载体技术快速扩增并向软骨分化人脂肪干细胞。[方法]将人脂肪干细胞结合Cytodex3微载体在旋转的生物反应器(RCSS)内进行动态培养,应用倒置显微镜和扫描电镜对微载体表面的脂肪干细胞进行动态观察,并对收获的脂肪干细胞进行Safran in-O、tolu id ine b lue染色等组织化学染色及Ⅱ型胶原的免疫化学染色分析。[结果]脂肪干细胞于24 h内贴附于Cytodex3微载体表面,细胞形态为短梭形,随时间的延长,贴附于微载体的细胞逐渐增多,到培养后期,细胞密度可达最初接种的19倍左右,在微载体上收获的细胞进行番红花O、阿利新蓝染色呈阳性,Ⅱ型胶原染色阳性,均强于对照组。[结论]利用微载体细胞培养技术可简便快速地在体外扩增脂肪干细胞,并成功实现向软骨细胞分化。
