Quality of Umbilical Cord Blood CD34+ Cells in a Double-Compartment Freezing Bag Cryopreserved without a Rate-Controlled Programmed Freezer
Quality of Umbilical Cord Blood CD34+ Cells in a Double-Compartment Freezing Bag Cryopreserved without a Rate-Controlled Programmed Freezer
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不使用速率控制程序冷冻机冷冻保存的双室冷冻袋中脐带血 CD34 细胞的质量
DOI：
10.1532/ijh97.06147
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发表时间：
2007
影响因子：
2.1
通讯作者：
S. Tsuchiya
中科院分区：
文献类型：
--
作者：
M. Minegishi;Tsuneo Itoh;Narumi Fukawa;T. Kitaura;J. Miura;Hiroyuki Takahashi;Akira Suzuki;Yoshinori Kudo;Ayuko Narita;Yuko Sato;Masakuni Suzuki;Takanori Watanabe;Yuichi Wada;Y. Takeyama;S. Tsuchiya
本研究的目的是评估一种简单的冷冻保存方法对脐带血CD34+细胞质量的影响。这些细胞被分配到一个双室冷冻袋中，在-85℃下冷冻保存，不使用速率控制式程序冷冻机，并保存在氮气的液态中。用7-氨基放线菌素D标记的流式细胞仪和集落形成实验检测冻融前后CD34+细胞的活性。分析20份超低温保存的脐血标本，平均保存时间为92天。冻存前CD34+细胞存活率为99.8%±0.4%，解冻后大室为99.5%±0.8%，小室为99.6%±0.5%，试管为99.4%±0.6%。冻存前活CD34+细胞集落形成实验的平均值分别为30.7±6.8(每100个活CD34+细胞的集落形成单位-粒细胞-巨噬细胞[CFU-GM])和68.5±14.8(每100个活CD34+细胞的总CFU)。解冻后的CFU-GM和总CFU分别为32.7±9.0和66.0±13.4，32.4±8.1和64.5±16.1，30.9±5.4和64.7±12.4。冷冻前和解冻后的集落形成试验结果无显著差异。总体而言，我们的发现表明，我们的简单方法在不使用速率控制式程序冷冻机的情况下冷冻保存脐血细胞，并不会损害脐血祖细胞的克隆形成能力。这种超低温保存方法可以为造血干细胞移植提供足够的细胞产品。
The aim of this study was to evaluate how a simple method of cryopreservation influences the quality of CD34+ cells in umbilical cord blood (UCB). The cells were dispensed into a double-compartment freezing bag, cryopreserved at -85°C without a rate-controlled programmed freezer, and stored in the liquid phase of nitrogen. The viability of the CD34+ cells before freezing and after thawing was assessed by flow cytometry with 7-aminoactinomycin D and by colony-forming assays. Twenty UCB units cryopreserved for a median of 92 days were analyzed. Mean CD34+ cell viabilities before freezing were 99.8% ± 0.4% and after thawing were 99.5% ± 0.8% in large chambers, 99.6% ± 0.5% in small chambers, and 99.4% ± 0.6% in sample tubes. The mean values from colony-forming assays of the viable CD34+ cells before freezing were 30.7 ± 6.8 (colony-forming units-granulocyte-macrophage [CFU-GM] per 100 viable CD34+ cells) and 68.5 ± 14.8 (total CFUs per 100 viable CD34+ cells). The CFU-GM and total CFU values after thawing were, respectively, 32.7 ± 9.0 and 66.0 ± 13.4 in large chambers, 32.4 ± 8.1 and 64.5 ± 16.1 in small chambers, and 30.9 ± 5.4 and 64.7 ± 12.4 in sample tubes. The results of the colony-forming assays before freezing and after thawing were not significantly different. Our findings overall indicated that our simple method for the cryopreservation of UCB cells without a rate-controlled programmed freezer does not impair the clonogenic capacity of UCB progenitor cells. This cryopreservation method could provide cellular products adequate for hematopoietic stem cell transplantation.
DOI：
10.1073/pnas.92.22.10119
发表时间：
1995-10-24
影响因子：
11.1
作者：
RUBINSTEIN, P;DOBRILA, L;STEVENS, CE
通讯作者：
STEVENS, CE
影响因子：
20.3
作者：
Wagner, JE;Barker, JN;Davies, SM
通讯作者：
Davies, SM
同时流式细胞术测量活力和淋巴细胞亚群增殖。
影响因子：
2.2
作者：
Schmid,I;Hausner,MA;Cole,SW;Uittenbogaart,CH;Giorgi,JV;Jamieson,BD
通讯作者：
Jamieson,BD