Martinson J A, Loudovaris M, Smith S L, Bender J G, Vachula M, van Epps D E, Kaizer H, Ghalie R G, McLeod B C
Baxter Healthcare Corp. Immunotherapy Division, Round Lake, Illinois 60073, USA.
J Hematother. 1997 Feb;6(1):69-75. doi: 10.1089/scd.1.1997.6.69.
Human CD34+ cells purified from frozen mobilized peripheral blood apheresis products (n = 7) were studied immediately (freshly isolated) or refrozen and studied after > 30 days storage in liquid nitrogen (refrozen/thawed). The proliferation and differentiation of freshly isolated or refrozen/thawed CD34+ cells were examined after 10 days of serum-supplemented suspension culture with recombinant human hematopoietic growth factors. The proliferative capacity (fold increase) of the refrozen/thawed CD34+ cells (mean +/- SD, 54.3 +/- 34.3) was comparable to the freshly isolated CD34+ cell cultures (49.0 +/- 42.4). Two-color flow cytometry of the CD34+ cultured cell populations, fresh and refrozen/thawed, displayed typical patterns of neutrophil differentiation into CD15/CD11b neutrophil precursors. The colony-forming ability of freshly isolated and refrozen/thawed CD34+ cells showed no significant differences (p > 0.05) in the total number or type of colony-forming units (CFU-GM, CFU-M, BFU-E, CFU-GEMM) obtained. In addition, the cloning efficiencies of freshly isolated (19.5 +/- 7.6%) and refrozen/thawed CD34+ cells (21.9 +/- 12.7%) were comparable (p = 0.366). These data suggest that CD34+ cells enriched from frozen apheresis blood products can be either used immediately or stored in liquid nitrogen and thawed with minimal effect on their ability to proliferate and differentiate in liquid culture.
从冷冻的动员外周血单采产品中纯化的人CD34+细胞（n = 7），立即进行研究（新鲜分离），或重新冷冻，并在液氮中储存超过30天后解冻进行研究（重新冷冻/解冻）。在用重组人造血生长因子进行血清补充悬浮培养10天后，检测新鲜分离或重新冷冻/解冻的CD34+细胞的增殖和分化情况。重新冷冻/解冻的CD34+细胞的增殖能力（增加倍数）（平均值±标准差，54.3±34.3）与新鲜分离的CD34+细胞培养物（49.0±42.4）相当。对新鲜和重新冷冻/解冻的CD34+培养细胞群体进行双色流式细胞术分析，显示中性粒细胞分化为CD15/CD11b中性粒细胞前体的典型模式。新鲜分离和重新冷冻/解冻的CD34+细胞的集落形成能力在获得的集落形成单位（CFU-GM、CFU-M、BFU-E、CFU-GEMM）总数或类型上无显著差异（p>0.05）。此外，新鲜分离的（19.5±7.6%）和重新冷冻/解冻的CD34+细胞的克隆效率（21.9±12.7%）相当（p = 0.366）。这些数据表明，从冷冻单采血液制品中富集的CD34+细胞既可以立即使用，也可以储存在液氮中，解冻后对其在液体培养中的增殖和分化能力影响最小。
