Liu Kai-Yan, Dong Wen-Chuan, Wang Yi-Lan, Jiang Yong-Jun, Gao Zhi-Yong, Huang Ning-Wei, Lu Dao-Pei
People Hospital, Institute of Hematology, Peking University, Beijing 100044, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2004 Oct;12(5):670-3.
This study was aimed to search for effective cryoprotectants and freezing methods used in cord blood bank (CBB) for cryopreservation of cord blood hematopoietic stem cells. The non-programmed group using 8% final concentration of dimethyl sulfoxide (DMSO) and 5% final concentration hydroxyethyl starch (HES) (molecular weight 120,000) as protectants and group of conventional of programmed controller method using 10% DMSO only as cryoprotectant in cryopreservation of cord blood hematopoietic stem cells were compared. In each of the two groups, 15 cord blood units were used. In non-programmed group, cord blood units put in -80 degrees C refrigerator for 24 hours as a transitional step before deep-freezing in liquid nitrogen, when both of DMSO and HES had been added. The recoveries of the nuclear cells number, the yield of granulocyto-macrophage colony forming units (CFU-GM) and the cells viability in cord blood units before preservation and after thawing were tested for both methods. The results showed that no significant difference was found in above assays between two groups. The clinical application results also showed that hematopoietic engraftment rates after infusion were similar in both groups. It is concluded that the non-programmed method by -80 degrees C refrigerator as a transitional step and using the combined two protectants seems simple in operation and effective in clinical transplantation as well as the conventional programmed method.
本研究旨在寻找脐带血库(CBB)中用于冷冻保存脐带血造血干细胞的有效冷冻保护剂和冷冻方法。比较了在脐带血造血干细胞冷冻保存中,使用终浓度8%的二甲基亚砜(DMSO)和终浓度5%的羟乙基淀粉(HES,分子量120,000)作为保护剂的非程序组,以及使用仅10%DMSO作为冷冻保护剂的传统程序控制方法组。两组各使用15个脐带血单位。在非程序组中,当同时加入DMSO和HES后,将脐带血单位放入-80℃冰箱24小时作为过渡步骤,然后再在液氮中深度冷冻。对两种方法保存前和解冻后脐带血单位中的核细胞数量回收率、粒-巨噬细胞集落形成单位(CFU-GM)产量和细胞活力进行了检测。结果显示,两组上述检测指标均无显著差异。临床应用结果也表明,两组输注后的造血植入率相似。结论是,以-80℃冰箱作为过渡步骤并使用两种保护剂联合的非程序方法在操作上似乎简单,在临床移植中也与传统程序方法一样有效。