制备了叔胺改性甘草次酸[GA-N(CH3)2]修饰的海藻酸钠[ALG-GA-N(CH3)2],并在温敏性琼脂糖的辅助作用下,利用微流体技术获得了高通量、单分散且粒径可控的ALG-GA-N(CH3)2微凝胶.考察了Span 80含量、疏水配体取代度、样品浓度和水/油相流速对微液滴制备的影响.研究结果表明,叔胺基改性可显著改善甘草次酸的亲水性;在Span 80质量分数为2.0%,疏水配体取代度小于12%,样品浓度小于15mg/m L,水相流速为1.5 m L/h,油相流速为6 m L/h条件下,可获得高通量、单分散及粒径为200μm的适用于细胞包封培养的微凝胶球.同时提供了一种三维培养肝细胞的新方法,为其在组织工程中的应用奠定了基础.
In order to obtain microgel sphere containing hydrophobic ligands which can be used for 3D liver cell culture. Firstly,the sodium alginate( ALG) was modified with tertiary ammonia-glycyrrhetinic acid[GA-N( CH3)2]. Then,the high flux,monodisperse,size controllable microgel of GA-N( CH3)2modified ALG[ALG-GA-N( CH3)2]was prepared by the technology of microfluidics with the assistant of agarose. The influence of different preparation condition,such as concentration of Span 80,substitution degree of hydrophobic ligand,concentration of ALG-GA-N( CH3)2,velocity of water/oil phase,on microdroplet preparation were also studied. The results showed that the introduction of tertiary ammonia can significantly improve the hydrophility of GA. And the monodisperse microgel with 200 μm diameter,which could be used for cell envelope culture,could be obtained under the condition below: the mass fraction of Span 80 is 2.0%,hydrophobic ligand substitution degree is less than 12%,concentration of sample is less than 15 mg / m L,the velocity of water is 1. 5 m L / h and the velocity of oil is 6.0 m L / h. The microgel provides a new method for liver cell 3D culture and laid the foundation for its application in tissue engineering.