不同微囊冷冻保存HK-2细胞效果比较

The Comparison of Cryopreserved Effects of Different Type of Microcapsules on HK-2 Cell

观察不同类型微囊冻存细胞的效果,为微囊冻存选择合适的冷冻保护剂。应用高压脉冲静电液滴发生器制作液化的海藻酸钠-多聚赖氨酸-海藻酸钠(APA)微囊和海藻酸钠-钙胶珠包被人肾小管上皮细胞(HK-2);用二甲基亚砜和甘油作为冷冻保护剂冷冻未包被的HK-2细胞、APA微囊包被的HK-2细胞、胶珠包被的HK-2细胞;解冻后用MTT法测定细胞活性、考马斯亮蓝法测定蛋白质含量、Hochest33258荧光染料测定DNA含量。结果表明,解冻后液化微囊内HK-2细胞的细胞活性、蛋白质含量、DNA含量均显著高于胶珠内细胞(P<0.05),但与未包被组差异不显著(P>0.05)。DMSO作为冷冻保护剂,细胞活性及细胞蛋白质含量均高于甘油组(P<0.05),但两者在DNA含量上无差异(P>0.05)。液化微囊与DM-SO合用时,细胞活性及细胞蛋白质含量、DNA含量均高于液化微囊与甘油合用时(P<0.05)。液化微囊解冻后稳定性优于胶珠。液化微囊冷冻保存效果优于胶珠,DMSO适于微囊冷冻。

In order to compare the cryopreservational effects of different types of microcapsules on cells and select the proper cryoprotectants for microencapsulation, HK-2 cells were encapsulated by liquefied alginate-polylyssine-alginate microcapsules and alginate-calcium microbeads. DMSO and glycerol were used as the cryoprotectants to cryopreserve unencapsulation ceils and encapsulation cells. The defrozed cell＇s viability was evaluated by MTT,and the protein contents and DNA contents were determined by Coomassie brilliant blue staine and fluorochrome Hochest 33258. The viability, protein contents, DNA contents of liquefied microcapsule＇s cells were significantly higher than that of cells encapsulated by microbeads （P 〈0. 05）, but there were no significant differences between liquefied microcapsule＇s cells and unencapsulated cells （P 〉0.05）. The cell viability,protein contents of the cells in DMSO as cryoproteetants were significantly higher than that in glycrerol （P 〈0.05） ,but there were no significant differences between them on DNA contents （P 〉0.05）. The liquefied microcapsules coupled with DMSO to cryopreserve cells were better than that coupled with glycerol in the cell viability, protein contents,DNA contents. The liquefied microcapsules were more stable than microbeads after being defrozed. The cryopreserved effect of the liquefied microcapsules was better than that of microbeads,and DMSO was more proper for microcapsules than glycerol.