Katayama Y, Yano T, Bessho A, Deguchi S, Sunami K, Mahmut N, Shinagawa K, Omoto E, Makino S, Miyamoto T, Mizuno S, Fukuda T, Eto T, Fujisaki T, Ohno Y, Inaba S, Niho Y, Harada M
Second Department of Internal Medicine, Okayama University Medical School, Japan.
Bone Marrow Transplant. 1997 Feb;19(3):283-7. doi: 10.1038/sj.bmt.1700644.
A simplified method for cryopreservation at -80 degrees C of peripheral blood stem cells (PBSC) has been increasingly used for autologous PBSC transplantation in Japan. Although this method, using 6% hydroxyethyl starch (HES) and 5% dimethyl sulfoxide (DMSO) as a cryoprotectant without rate-controlled freezing, has several advantages over the conventional method using 10% DMSO with rate-controlled freezing, little is known about effects of long-term cryopreservation for years and thawing process on hematopoietic progenitors. We examined the recovery rates of BFU-E and CFU-GM in sample tubes cryopreserved by the simplified method under various conditions as follows: (1) long-term storage for 1-5 years; (2) DMSO exposure for 1 h after rapid thawing; and (3) thawing at a lower temperature other than 37 degrees C. In our study, we found that the recovery rates of BFU-E and CFU-GM were not affected by the length of cryopreservation period; they remained at more than 70% on average for 16-61 months. In our hands, a 1-h exposure to DMSO after rapid thawing was not toxic for hematopoietic progenitors. Furthermore, there was no significant difference in the recovery rates of BFU-E and CFU-GM between thawing at 37 degrees C and 20 degrees C. These observations indicate that PBSC cryopreserved for at least 5 years by the simplified method can be used clinically without losing hematopoietic activity, and suggest that hematopoietic activity of the thawed PBSC may be unaffected when PBSC are infused slowly within 60 min or even when PBSC are thawed gradually at room temperature.
在日本，一种用于外周血干细胞（PBSC）在-80℃下冷冻保存的简化方法已越来越多地用于自体PBSC移植。尽管这种方法使用6%羟乙基淀粉（HES）和5%二甲基亚砜（DMSO）作为冷冻保护剂且无速率控制冷冻，与使用10% DMSO并进行速率控制冷冻的传统方法相比有几个优点，但对于多年的长期冷冻保存和 thawing 过程对造血祖细胞的影响知之甚少。我们在以下各种条件下检查了通过简化方法冷冻保存的样品管中BFU-E和CFU-GM的回收率：（1）长期保存1至5年；（2）快速解冻后DMSO暴露1小时；（3）在37℃以外的较低温度下解冻。在我们的研究中，我们发现BFU-E和CFU-GM的回收率不受冷冻保存期长度的影响；在16至61个月内平均保持在70%以上。在我们的操作中，快速解冻后1小时的DMSO暴露对造血祖细胞无毒。此外，在37℃和解冻在20℃之间，BFU-E和CFU-GM的回收率没有显著差异。这些观察结果表明，通过简化方法冷冻保存至少5年的PBSC可在临床上使用而不丧失造血活性，并表明当PBSC在60分钟内缓慢输注甚至当PBSC在室温下逐渐解冻时，解冻后的PBSC的造血活性可能不受影响。
