CAJ | 학술논문

目的探讨-80℃非程控冷冻保存人外周血干细胞(PBSC)的效果.方法 41份PBSC标本,39份采自恶性肿瘤患者,2份采自正常供者.对于恶性肿瘤患者,干细胞采集前采用针对性化疗方案联合粒细胞集落刺激因子或粒-单细胞集落刺激因子进行动员.将采集的干细胞与采用120 g/L羟乙基淀粉、体积分数为10%二甲基亚砜及80 g/L人血清白蛋白组成的冷冻保护剂按1:1混合,然后将其直接放入-80℃低温冰箱内保存.测定保存前以及保存后不同时间的单个核细胞(MNC)、有核细胞(NC)、CD34+细胞、粒一单核细胞系集落形成单位(CFU-GM)和红系爆式集落形成单位(BFU-E)等的回收率以及细胞的台盼蓝拒染率.结果经-80℃非程控直接冷冻保存1个月至10年,PBSC的台盼蓝拒染率、NC和MNC回收率的变化较小(P>0.05),而CD34+细胞、CFU-GM和BFU-E回收率在冻存5～10年后下降较明显(P＜0.05),但仍分别达89.6%、85.1%和83.7%.34例患者经相应处理后,回输经-80℃下保存13～53 d(平均19 d)的干细胞,患者于细胞回输后11～27 d(平均14.7 d)获得造血功能重建.结论在采用终浓度为60 g/L 羟乙基淀粉、体积分数为5%二甲基亚砜及40 g/L人血清白蛋白组成的冷冻保护剂的情况下,-80℃非程控冷冻法适于PBSC的长期保存,效果较理想.
목적 탐토-80℃비정공냉동보존인외주혈간세포(PBSC)적효과.방법 41빈PBSC표본,39빈채자악성종류환자,2빈채자정상공자.대우악성종류환자,간세포채집전채용침대성화료방안연합립세포집락자격인자혹립-단세포집락자격인자진행동원.장채집적간세포여채용120 g/L간을기정분、체적분수위10%이갑기아풍급80 g/L인혈청백단백조성적냉동보호제안1:1혼합,연후장기직접방입-80℃저온빙상내보존.측정보존전이급보존후불동시간적단개핵세포(MNC)、유핵세포(NC)、CD34+세포、립일단핵세포계집락형성단위(CFU-GM)화홍계폭식집락형성단위(BFU-E)등적회수솔이급세포적태반람거염솔.결과 경-80℃비정공직접냉동보존1개월지10년,PBSC적태반람거염솔、NC화MNC회수솔적변화교소(P>0.05),이CD34+세포、CFU-GM화BFU-E회수솔재동존5～10년후하강교명현(P＜0.05),단잉분별체89.6%、85.1%화83.7%.34례환자경상응처리후,회수경-80℃하보존13～53 d(평균19 d)적간세포,환자우세포회수후11～27 d(평균14.7 d)획득조혈공능중건.결론 재채용종농도위60 g/L 간을기정분、체적분수위5%이갑기아풍급40 g/L인혈청백단백조성적냉동보호제적정황하,-80℃비정공냉동법괄우PBSC적장기보존,효과교이상.
Objective To investigate the cryopreservation of human peripheraI blood stern cells (PBSC)by-80℃non rate-controlled freezing method.Methods Forty-one samples of PBSC were obtained from 39 malignant tumor patients and 2 nornlal donors.PBSC in all malignant tumor patients were mobilized with special cherootherapy protocols combined with granulocyte colony-stimulating factor or granulocyte/monocyte colony-stimulating factor before collection.The obtained PBSC were mixed with same volume of cryoprotectant composed of 12 g/L hydroxyethyl starch,10% dimethylsulfoxide and 80 g/L human serunl albumin,and then eryopreserved in-80℃deep freeze refrigerator directly.Trypan blue viability and recovery rate of nucleated cells(NC),mononuclear cells(MNC),CD34+cells(CD34+),colony-forming unit-granulocyte macrophages (CFU-GM),burst forming unit-erythroid(BFU-E) were detected before and after cryopreservation in different periods.Results Having been cryopreserved from 1 month to 10 years in-80℃.there were no statistically significant differences in trypan blue viability,NC and MNC recovery rate(P>0.05).AIthough recovery rate of CD34+, CFU-M and BFU-E was decreased obviously after cryopreservation for 5-10 years(P＜0.05),10-year recovery rate was 89.6%,85.1%and 83.7%,respectively.Thirty-four patients subject to corresponding pretreatment were transfused with-80℃ cryopreserved PBSC which had been stored for 13 to 35 days(mean 19 days)and hematological reconstitution was obtained successfully in 11 to 27 days (mean 14.7 days).Conclusion -80℃non rate-controlled freezing method is suitable to human PBSC long-term cryopreservation with the cryoproteetant composed of 60 g/L hydroxyethyl starch,5%dimethylsulfoxide and 40 g/L human serum albumin,and its cryopreservation effect is ideal