Muramaki Mototsugu, Hara Isao, Miyake Hideaki, Yamada Yuji, Okada Hiroshi, Kamidono Sadao
Department of Organ Therapeutics, Faculty of Medicine, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.
Oncol Rep. 2003 Nov-Dec;10(6):1993-8.
The objectives of this study were to evaluate the hematopoietic activity of peripheral blood stem cells (PBSCs) harvested from patients with advanced germ cell tumors after long-term cryopreservation in a mechanical freezer at -80 degrees C. Forty-one patients with advanced tumors were enrolled in this study. Flow cytometric analysis was performed to determine the total cell viability and the viability of CD34 positive cells of PBSCs stored between January 1995 and December 2001. The 41 patients were divided into two groups according to the duration of cryopreservation. In the 21 patients in whom the duration of storage was greater than 5 years, the total cell number did not change significantly before and after long-term cryopreservation (p=0.75). The percentage of viable cells and the total number of viable cells was significantly decreased after long-term cryopreservation (p<0.001 and p<0.05, respectively). However, the percentage of CD34 positive cells was significantly increased to 2.34% from 1.85% (p<0.05), resulting in non-statistically significant change in the total viable CD34 positive cells (p=0.98). A similar relation was found in the 20 patients whose storage duration was less than 2 years, although a significant change was found only in the percentage of viable cells. Moreover, pre-PBSCH factors, including age, cumulative dosage of cisplatin and the International Germ Cell Cancer Collaborative Group classification at the diagnosis did not affect the viable CD34 positive cell count after long-term cryopreservation. In conclusion, the present results suggest that CD34 positive cells could be successfully cryopreserved in a mechanical freezer at -80 degrees C for more than 5 years with no relation to pre-PBSCH factors and efficacy of PBSCH itself.
本研究的目的是评估晚期生殖细胞肿瘤患者外周血干细胞（PBSCs）在-80℃机械冷冻箱中长期冷冻保存后的造血活性。41例晚期肿瘤患者纳入本研究。对1995年1月至2001年12月期间保存的PBSCs进行流式细胞术分析，以确定总细胞活力和CD34阳性细胞的活力。根据冷冻保存时间将41例患者分为两组。在保存时间超过5年的21例患者中，长期冷冻保存前后总细胞数无显著变化（p=0.75）。长期冷冻保存后活细胞百分比和活细胞总数显著降低（分别为p<0.001和p<0.05）。然而，CD34阳性细胞百分比从1.85%显著增加至2.34%（p<0.05），导致存活CD34阳性细胞总数无统计学显著变化（p=0.98）。在保存时间少于2年的20例患者中也发现了类似关系，尽管仅在活细胞百分比方面有显著变化。此外，PBSCH前因素，包括年龄、顺铂累积剂量和诊断时的国际生殖细胞癌协作组分类，均不影响长期冷冻保存后存活CD34阳性细胞计数。总之，目前的结果表明，CD34阳性细胞可在-80℃机械冷冻箱中成功冷冻保存5年以上，且与PBSCH前因素及PBSCH本身的疗效无关。
