Rodríguez Luciano, Velasco Beatriz, García Joan, Martín-Henao Gregorio Angel
Transfusion Center and Tissue Bank, Cell Therapy Unit, Hospital Duran i Reynals, Barcelona, Spain.
Transfusion. 2005 Aug;45(8):1391-7. doi: 10.1111/j.1537-2995.2005.00213.x.
The direct transfusion of thawed hematopoietic progenitor cells (HPCs) is associated to transfusion-related side effects that are thought to be dose-dependent on the infused dimethyl sulfoxide (DMSO). Both the effectiveness of a fully automated cell processing device to washing out DMSO and the effects of DMSO elimination over the recovered cells were evaluated.
Twenty cryopre-served peripheral blood HPC bags (HPC apheresis [HPC-A]) were thawed and processed for washing with an automated cell-processing device. Viability, colony-forming units (CFUs), and absolute count of recovered cells were evaluated by flow cytometry immediately after washing as well as at different times after washing and compared with a sample taken just after thawing (control) but maintained at 4 degrees C. DMSO content was measured by high-performance liquid chromatography and the osmolarity with an osmometer.
The median recovery of viable total nucleated cells, viable CD34+ cells, and CFU colonies was 89 (range, 74-115), 103 (range, 62-126), and 91 percent (range, 46%-196%), respectively, in the washing group. Recovery of viable CD3+ cells was 97 percent (range, 42%-131%) and CD14+ cells was 82 percent (54%-119%). The percentages of DMSO elimination and osmolarity reduction were 98 (range, 96-99) and 90 percent (range 86%-95%), respectively. Moreover, elimination of the cryoprotectant improved CFU count, viability, and cell recoveries along the time when compared with the control group.
Washing out DMSO in thawed HPC-A by use of this approach is safe and efficient in terms of recovery and viability of nucleated and progenitor cells. Additionally, the removal degree of DMSO is very high and therefore might ameliorate the transfusion-related side effects.
输注解冻后的造血祖细胞（HPC）与输血相关的副作用有关，这些副作用被认为与输注的二甲基亚砜（DMSO）剂量相关。本研究评估了全自动细胞处理设备清除DMSO的效果以及DMSO清除对回收细胞的影响。
将20袋冷冻保存的外周血HPC（单采外周血造血干细胞[HPC-A]）解冻，并用全自动细胞处理设备进行洗涤处理。洗涤后立即以及洗涤后的不同时间，通过流式细胞术评估回收细胞的活力、集落形成单位（CFU）和绝对计数，并与解冻后立即采集但保存在4℃的样本（对照组）进行比较。通过高效液相色谱法测量DMSO含量，并用渗透压计测量渗透压。
洗涤组中，有核活细胞、CD34+活细胞和CFU集落的中位回收率分别为89%（范围74%-115%）、103%（范围62%-126%）和91%（范围46%-196%）。CD3+活细胞的回收率为97%（范围42%-131%），CD14+活细胞的回收率为82%（54%-119%）。DMSO清除率和渗透压降低率分别为98%（范围96%-99%）和90%（范围86%-95%）。此外，与对照组相比，随着时间推移，冷冻保护剂的清除改善了CFU计数、活力和细胞回收率。
使用这种方法洗涤解冻后的HPC-A中的DMSO，在有核细胞和祖细胞的回收率和活力方面是安全有效的。此外，DMSO的去除程度非常高，因此可能会改善输血相关的副作用。
