开放大孔海藻酸微球的制备及三维细胞培养应用  

Preparation of Open Macroporous Alginate Microspheres for Three-Dimensional Cell Culture

作　　者：陈思珂 巨晓洁[1] Sike Chen;Xiaojie Ju(School of Chemical Engineering,Sichuan University,Chengdu 610065,China)

机构地区：[1]四川大学化学工程学院,四川成都610065

出　　处：《高分子材料科学与工程》2023年第8期126-133,共8页Polymer Materials Science & Engineering

基　　金：国家自然科学基金资助项目(22078202)。

摘　　要：开放大孔微球能够帮助细胞-细胞和细胞-细胞外基质相互作用的形成,适合作为三维细胞培养支架。采用微流控技术和在线紫外引发交联法制备得到尺寸均一、大小可控的甲基丙烯酰化海藻酸(AlgMA)微球,利用冷冻干燥制孔法得到具有开放大孔的AlgMA微球。系统研究了内/外相流速比的变化和AlgMA浓度对微球尺寸、形貌和稳定性的影响,并以人肝癌细胞HepG2作为模型细胞,系统考察了AlgMA微球对细胞活性和增殖的影响。结果表明,当Alg-MA质量分数为10%时,微球的复溶胀性能和孔洞结构最好,分别培养HepG2细胞24 h,48 h和72 h后,细胞活性均在85%以上,培养48 h和72 h后,相较于前一天细胞数量分别倍增了1.24倍和1.76倍。该研究工作为开放大孔海藻酸微球的制备提供了新方法。Open macroporous microspheres can assist in the formation of cell-cell and cell-extracellular matrix interactions which are suitable as three-dimensional cell culture scaffolds.Uniform and controllable sized methacrylated alginate(AlgMA)microspheres were prepared by microfluidic technology and online UV crosslinking method.AlgMA microspheres with open macropores were obtained by freeze-drying method.The changes in internal/external phase flow rate ratio and effect of AlgMA concentration on the size,morphology,and stability of microspheres were systematic studied.Using human liver cancer cell HepG2 as model cell,the effects of AlgMA microspheres on cell activity and proliferation were systematically investigated.The results show that when the mass fraction of AlgMA is 10%,the reswelling performance and pore structure of the microspheres are the best.After 24 h,48 h,and 72 h of culture,the cell activity of HepG2 cells is above 85%.After 48 h and 72 h of cultivation,the numbers of cells double by 1.24 and 1.76 times compared to the previous day,respectively.This research work provides a new method for the preparation of open connected macroporous alginate microspheres.

关 键 词：开放大孔微球 海藻酸微球 三维细胞培养 甲基丙烯酰化海藻酸 肝癌细胞 

分 类 号：Q813.11[生物学—生物工程]