Reddy Opal L, Sall Mame Thioye, Dinh Anh, Cai Yihua, Ongkeko Martin, Arya Nina, Wilder Jennifer, Tran Minh, Jin Ping, Stroncek David F, Panch Sandhya R
Department of Pathology, Keck School of Medicine of USC, Los Angeles, California, USA.
Center for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, Maryland, USA.
Transfusion. 2023 Apr;63(4):774-781. doi: 10.1111/trf.17314. Epub 2023 Mar 28.
Since the beginning of the COVID-19 pandemic, cryopreservation of hematopoietic progenitor cell (HPC) products has been increasingly used to ensure allogeneic donor graft availability prior to recipient conditioning for transplantation. However, in addition to variables such as graft transport duration and storage conditions, the cryopreservation process itself may adversely affect graft quality. Furthermore, the optimal methods to assess graft quality have not yet been determined.
A retrospective review was performed on all cryopreserved HPCs processed and thawed at our facility from 2007 to 2020, including both those collected onsite and by the National Marrow Donor Program (NMDP). HPC viability studies were also performed on fresh products, retention vials, and corresponding final thawed products by staining for 7-AAD (flow cytometry), AO/PI (Cellometer), and trypan blue (manual microscopy). Comparisons were made using the Mann-Whitney test.
For HPC products collected by apheresis (HPC(A)), pre-cryopreservation and post-thaw viabilities, as well as total nucleated cell recoveries were lower for products collected by the NMDP compared to those collected onsite. However, there were no differences seen in CD34+ cell recoveries. Greater variation in viability testing was observed using image-based assays compared to flow-based assays, and on cryo-thawed versus fresh samples. No significant differences were observed between viability measurements obtained on retention vials versus corresponding final thawed product bags.
Our studies suggest extended transport may contribute to lower post-thaw viabilities, but without affecting CD34+ cell recoveries. To assess HPC viability prior to thaw, testing of retention vials offers predictive utility, particularly when automated analyzers are used.
自新冠疫情开始以来,造血祖细胞(HPC)产品的冷冻保存越来越多地用于在受体移植预处理之前确保异基因供体移植物的可用性。然而,除了移植物运输时间和储存条件等变量外,冷冻保存过程本身可能会对移植物质量产生不利影响。此外,评估移植物质量的最佳方法尚未确定。
对2007年至2020年在我们机构处理和解冻的所有冷冻保存的HPC进行回顾性研究,包括现场采集的和通过国家骨髓捐赠计划(NMDP)采集的。还通过7-AAD染色(流式细胞术)、AO/PI染色(细胞计数仪)和台盼蓝染色(手动显微镜检查)对新鲜产品、留存小瓶和相应的最终解冻产品进行了HPC活力研究。使用曼-惠特尼检验进行比较。
对于通过单采术采集的HPC产品(HPC(A)),与现场采集的产品相比,NMDP采集的产品在冷冻保存前和解冻后的活力以及总核细胞回收率较低。然而,CD34+细胞回收率没有差异。与基于流式细胞术的检测方法相比,基于图像的检测方法在冷冻解冻样本与新鲜样本上观察到更大的活力检测差异。在留存小瓶与相应的最终解冻产品袋上获得的活力测量值之间未观察到显著差异。
我们的研究表明,延长运输可能导致解冻后活力降低,但不影响CD34+细胞回收率。为了在解冻前评估HPC活力,对留存小瓶进行检测具有预测作用,特别是在使用自动分析仪时。