γ-聚谷氨酸在细胞冻存中的作用  

Cryoprotective Effect of γ-Polyglutamic Acid on Cells

作　　者：陈焕芸 应丹妮 蔡海波[1] 谭文松[1] CHEN Huan-yun;YING Dan-ni;CAI Hai-bo;TAN Wen-song(State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《高分子通报》2020年第4期30-36,共7页Polymer Bulletin

摘　　要：无DMSO、无血清细胞冻存液在临床级细胞产品的冷冻保存中具有重要的应用价值。本文以K562细胞为模型,设置含10%(v/v)DMSO及胎牛血清的冻存液为阳性对照,含10%(v/v)甘油的RPMI 1640培养基冻存液为阴性对照组,含10%(v/v)甘油及0.01%(w/v)γ-PGA的RPMI 1640培养基冻存液为实验组,考察了在无DMSO、无血清细胞冻存体系中γ-聚谷氨酸对细胞的冷冻保护作用。将K562细胞以1×10^6cells/mL的密度分别悬浮在上述冻存液中,置于液氮冻存10周,检测复苏后K562的细胞复苏率、细胞形态以及复苏后细胞的扩增情况,以评判γ-聚谷氨酸在无DMSO无血清冻存液中对K562细胞的冷冻保护作用。结果显示,实验组的细胞复苏率为(83.00±3.00)%,明显高于阴性对照组的(70.33±5.51)%(p<0.05)和阳性对照组的(71.00±2.65)%(p<0.05);且实验组冻存后的细胞形态完整,冻存前后细胞的平均直径及圆度基本一致,细胞复苏后培养24h后的细胞活性为(88.83±14.29)%,明显高于阴性对照组的(67.51±5.20)%(p<0.05),与阳性对照组的(78.75±3.31)%没有显著性差异,同时复苏后细胞扩增的延滞期明显缩短。可见,在无DMSO、无血清的甘油冻存体系中添加γ-PGA可显著提高细胞的冻存效果,具有良好的实际应用价值。DMSO-free and Serum-free cell freezing medium has important application value in the cryopreservation of clinical grade cell products.In this paper,K562 cells were used as model cells to explore the cryoprotective effect of γ-PGA on cells in a DMSO-free and Serum-free system.The RPMI 1640 medium with glycerol content of 10%(v/v)was set as the negative control group,the fetal bovine serum with DMSO content of 10%(v/v)was set as the positive control group,the RPMI 1640 medium with glycerol content of 10%(v/v)andγ-PGA content of 0.01%(w/v)was set as the experimental group.K562 cells were suspended in the above cell freezing medium at a density of 1×10^6 cells/mL and placed in liquid nitrogen for 10 weeks.After resuscitation,cell recovery rate,cell morphology,and cell expansion ability of K562 were detected to analyze the cryoprotective effect ofγ-polyglutamic acid on K562 cells in DMSO-Free and Serum-Free cell freezing medium.The results showed that the cell recovery rate of the experimental group was(83.00±3.00)%,which was significantly higher than that of the negative control group(70.33±5.51)%(p<0.05)and the positive control group(71.00±2.65)%(p<0.05);the morphology of the cells in the experimental group was intact after freezing,the average diameter and roundness of cells were basically the same before and after freezing;the cell viability after culture for 24 h after resuscitation was(88.83±14.29)%,which was significantly higher than that of the negative control group(67.51±5.20)%(p<0.05),and was not significantly different from the(78.75±3.31)%of the positive control group;the lag phase in cell expansion after resuscitation was significantly shortened.It can be seen that the addition of γ-PGA to the glycerol cryopreservation system without DMSO and serum can significantly improve the cryopreservation effect of cells,and has good practical application value.

关 键 词：低温保存 聚谷氨酸 无DMSO细胞冻存液 无血清细胞冻存液 

分 类 号：R318.52[医药卫生—生物医学工程]