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

张丽娟; 秦雨琪; 任伟东; 董坚

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

昆明医科大学第三附属医院

基金项目:

基金：云南省科技厅基金资助项目 (2015DG034);

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出版历程
- 收稿日期: 2017-02-14

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

Funds:

基金：云南省科技厅基金资助项目 (2015DG034);

摘要

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.
- 冻存保护剂 /
- DMS /
- 人血白蛋白 /
- 勃脉力A /
- 安全性
Abstract: Objective To investigate a safe and effective clinical-grade cryoprotectant, which can be infused to patients. Me thods We collected 50 samples 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.Re s ults 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. Conclus ion The cryoprotectants with 7.5% DMSO +10% human serum albumin+ Plasmalyte A is safe and effective for cryopreserving the nucleated cells.
- Cryoprotectant /
- DMSO /
- Human serum albumin /
- Plasmalyte A /
- Safety

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参考文献(22)

[1]	[1]VALERI C R, PIVACEK L E.Effects of the temperature, the duration of frozen storage, and the freezing container on in vitro measurements in human peripheral blood mononuclear cells[J].Transfusion, 1996, 36 (4) :303-308.
[2]	[2]CILLONI D, GARAU D, REGAZZI E, et al.Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing[J].Bone marrow transplantation, 1999, 23 (5) :497-503.
[3]	[3]KATAYAMA Y, YANO T, BESSHO A, et al.The effects of a simplified method for cryopreservation and thawing procedures on peripheral blood stem cells[J].Bone Marrow Transplant, 1997, 19 (3) :283-287.
[4]	[4]CORDOBA R, ARRIETA R, KERGUELEN A, et al.The occurrence of adverse events during the infusion of autologous peripheral blood stem cells is related to the number of granulocytes in the leukapheresis product[J].Bone marrow transplant, 2007, 40 (11) :1063-1067.
[5]	[5]COX M A, KASTRUP J, HRUBIKO M.Historical perspectives and the future of adverse reactions associated with haemopoietic stem cells cryopreserved with dimethyl sulfoxide[J].Cell Tissue Bank, 2012, 13 (2) :203-215.
[6]	[6]SHU Z, HEIMFELD S, GAO D.Hematopoietic Stem Cell Transplantation with Cryopreserved Grafts:Adverse Reactions after Transplantation and Cryoprotectant Removal Prior to Infusion[J].Bone marrow transplantation, 2014, 49 (2) :469.
[7]	[7]HIGMAN MA, PORT J D, BEAUCHAMP NJ JR, et al.Reversible leukoencephalopathy associated with re-infusion of DMSO preserved stem cells[J].Bone Marrow Transplant, 2000, 26 (7) :797-800.
[8]	[8]KEUNG Y K, LAU S, ELKAYAM U, et al.Cardiac arrhythmia after infusion of cryopreserved stem cells[J].Bone Marrow Transplant, 1994, 14 (3) :363-367.
[9]	[9]LPEZ-JIMNEZ J, CERVER C, MUOZ A, et al.Cardiovascular toxicities related to the infusion of cryopreserved grafts:results of a controlled study[J].Bone Marrow Transplant, 1994, 13 (6) :789-793.
[10]	[10]RODRGUEZ L, VELASCO B, GARC A J, et al.Evaluation of an automated cell processing device to reduce the dimethyl sulfoxide from hematopoietic grafts after thawing[J].Transfusion, 2005, 45 (8) :1391-1397.
[11]	[11]FOS E, DESMARTIN M, BENHAMIDA S, et al.Recovery, viability and clinical toxicity of thawed and washed haematopoietic progenitor cells:analysis of 952 autologous peripheral blood stem cell transplantations[J].Bone Marrow Transplant, 2007, 40 (9) :831-835.
[12]	[12]AKK K A, HOLTE MR, TANGEN J M, et al.Hematopoietic engraftment of dimethyl sulfoxide depleted autologous peripheral blood progenitor cells[J].Transfusion, 2009, 49 (2) :354-361.
[13]	[13]MERYMAN H T.Cryoprotective agents[J].Cryobiology, 1971, 8 (2) :173-183.
[14]付旭锋, 司维, 郑冰蓉, 等.间充质干细胞冻存方法的研究进展[J].云南大学学报 (自然科学版) , 2016, 38 (4) :652-660.
[15]张苗, 张伯龙, 张佐云, 等.一种简便的外周血干细胞冷冻保存法[J].中国实验血液学杂志, 2001, 9 (4) :363-367.
[16]	[16]ABRAHAMSEN J F, BAKKEN A M, BRUSERUD, et al.Cryopreserving human peripheral blood progenitor cells with 5 percent rather than 10-percent DMSO results in less apoptosis and necrosis in CD34+cells[J].Transfusion, 2002, 42 (12) :1573-1580.
[17]	[17]ABRAHAMSEN J F, RUSTEN L, BAKKEN A M, et al.Better preservation of early hematopoietic progenitor cells when human peripheral blood progenitor cells are cryopreserved with 5 percent dimethylsulfoxide instead of 10 percent dimethylsulfoxide[J].Transfusion, 2004, 44 (5) :785-789.
[18]	[18]BAKKEN A, BRUSERUD O, ABRAHAMSEN J, et al.No differences in colony formation of peripheral blood stem cells frozen with 5%or 10%dimethyl sulfoxide.Journal of hematotherapy&stem cell research[J].Journal of hematotherapy&stem cell research, 2003, 12 (3) :351-358.
[19]	[19]VEERAPUTHIRAN M, THEUS JW, PESEK G, et al.Viability and engraftment of hematopoietic progenitor cells after long-term cryopreservation:effect of diagnosis and percentage dimethyl sulfoxide concentration[J].Cytotherapy, 2010, 12 (6) :764-766.
[20]娄琳, 陆志刚.海藻糖深低温保存外周血造血干细胞的可行性[J].中国组织工程研究与临床康复.2010, 14 (12) :2119-2122.
[21]	[21]SMAGUR A, MITRUS I, GIEBEL S, et al.Impact of different dimethyl sulphoxide concentrations on cell recovery, viability and clonogenic potential of cryopreserved peripheral blood hematopoietic stem and progenitor cells[J].Vox Sanguinis, 2013, 104 (3) :240-247.
[22]	[22]MITRUS I, SMAGUR A, GIEBEL S, et al.A faster reconstitution of hematopoiesis after autologous transplantation of hematopoietic cells cryopreserved in 7.5%dimethyl sulfoxide if compared to 10%dimethyl sulfoxide containing medium[J].Cryobiology, 2013, 67 (3) :327-331.

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