直接用于输注的临床级细胞冻存保护剂的动物体内外安全性评价  

In Vivo and In Vitro Safety Evaluation of Clinical-grade Cryoprotectant

作　　者：张丽娟[1] 秦雨琪 任伟东[1] 董坚[1] ZHANG Li-juan;QIN Yu-qi;REN Wei-dong;DONG Jian(The 3rd Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650101,China)

机构地区：[1]昆明医科大学第三附属医院,云南昆明650101

出　　处：《昆明医科大学学报》2017年第6期19-24,共6页Journal of Kunming Medical University

基　　金：云南省科技厅基金资助项目(2015DG034)

摘　　要：目的探索研究一种安全有效的可以直接用于输注的临床级细胞冻存保护剂.方法采集50份脐带血,使用羟乙基淀粉沉淀法分离得到脐血有核细胞,加入5种不同浓度的冻存保护剂(DMSO、人血白蛋白、勃脉力A),于液氮中冻存6个月,检测并比较冻存前后有核细胞、CD34+细胞、细胞集落形成单位、回收率和台盼蓝拒染率;将不同浓度的冻存保护剂经尾静脉注射到小鼠体内,评估其安全性.结果 A组(10%DMSO+5%人血白蛋白+勃脉力A)与C组(7.5%DMSO+10%人血白蛋白+勃脉力A)台盼蓝拒染率、有核细胞回收率、CD34+细胞回收率和集落形成单位回收率无统计学差异(P>0.05)且高于其它3组(P<0.05),但A组的肝脏毒性高于其它组(P<0.05).结论 7.5%DMSO+10%人血白蛋白+勃脉力A是一种安全有效的冻存保护剂.Objective To investigate a safe and effective clinical-grade cryoprotectant,which can be infused to patients.Methods We collected50samples of cord blood using hydroxyethyl starch precipitation method to get cord blood nucleated cells.Then we added five kinds of different concentration cryoprectants including DMSO,human serum albumin and Plasmalyte A to cryopreserve nucleated cells in liquid nitrogen.The effects of these cryoprotectants on biological properties of cord blood nucleated cells were monitored and compared after the cells were refrigerated for six months.The detected biological properties included the number of total nucleated cells,CD34+cells,colony forming units,recovery rate and trypan blue rejection rate.Cryoprotectants of different concentrations were injected into the mice of each group via tail vein,and the liver and renal toxicity were evaluated.Results There was no difference between group A and group C which resulted in higher recovery rates of nucleated cells,CD34+cells and colony forming units and trypan blue rejection rate than other three groups.The liver toxicity of group A were higher than other groups.Conclusion The cryoprotectants with7.5%DMSO+10%human serum albumin+Plasmalyte A is safe and effective for cryopreserving the nucleated cells.

关 键 词：冻存保护剂 DMS 人血白蛋白 勃脉力A 安全性 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]