Eichler Hermann, Kern Susanne, Beck Christian, Zieger Wolfgang, Klüter Harald
Institute of Transfusion Medicine and Immunology, Red Cross Blood Service of Baden-Württemberg-Hessen, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Germany.
Stem Cells. 2003;21(2):208-16. doi: 10.1634/stemcells.21-2-208.
Umbilical cord blood (UCB) preparation needs to be optimized in order to develop more simplified procedures for volume reduction, as well as to reduce the amount of contaminating cells within the final stem cell transplant. We evaluated a novel filter device (StemQuick((TM))E) and compared it with our routine buffy coat (BC) preparation procedure for the enrichment of hematopoietic progenitor cells (HPCs). Two groups of single or pooled UCB units were filtered (each n = 6), or equally divided in two halves and processed by filtration and BC preparation in parallel (n = 10). The engraftment capacity of UCB samples processed by whole blood (WB) preparation was compared with paired samples processed by filtration in the nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse animal model. Filtration of UCB units in the two groups with a mean volume of 87.8 and 120.7 ml, respectively, and nucleated cell (NC) content of 9.7 and 23.8 x 10(8) resulted in a sufficient mean cell recovery for mononucleated cells ([MNCs] 74.2%-77.5%), CD34(+) cells (76.3%-79.0%), and colony-forming cells (64.1%-86.3%). Moreover, we detected a relevant depletion of the transplants for RBCs (89.2%-90.0%) and platelets ([PLTs] 77.5%-86.1%). In contrast, the mean depletion rate using BC processing proved to be significantly different for PLTs (10%, p = 0.03) and RBCs (39.6%, p < 0.01). The NC composition showed a highly significant increase in MNCs and a decrease in granulocytes after filtration (p < 0.01), compared with a less significant MNC increase in the BC group (p < 0.05). For mice transplanted with WB-derived progenitors, we observed a mean of 15.3% +/- 15.5% of human CD45(+) cells within the BM compared with 19.9% +/- 16.8% for mice transplanted with filter samples (p = 0.03). The mean percentage of human CD34(+) cells was 4.2% +/- 3.1% for WB samples and 4.5% +/- 3.2% for filter samples (p = 0.68). As the data of NOD/SCID mice transplantation demonstrated a significant engraftment capacity of HPCs processed by filtration, no negative effect on the engraftment potential of filtered UCB cells versus non-volume-reduced cells from WB transplants was found. The StemQuick((TM))E filter devices proved to be a useful tool for Good Manufacturing Practices conform enrichment of HPCs and MNCs out of UCB. Filtration enables a quick and standardized preparation of a volume-reduced UCB transplant, including a partial depletion of granulocytes, RBCs, and PLTs without the need for centrifugation. Therefore, it seems very probable that filter-processed UCB transplants will also result in sufficient hematopoietic reconstitution in humans.
为了开发更简化的体积减少程序，并减少最终干细胞移植中污染细胞的数量，脐带血（UCB）制备需要进行优化。我们评估了一种新型过滤装置（StemQuick((TM))E），并将其与我们常规的用于富集造血祖细胞（HPC）的血沉棕黄层（BC）制备程序进行了比较。两组单份或汇集的UCB样本进行了过滤（每组n = 6），或均分为两半并分别通过过滤和BC制备并行处理（n = 10）。在非肥胖糖尿病/严重联合免疫缺陷（NOD/SCID）小鼠动物模型中，将通过全血（WB）制备处理的UCB样本的植入能力与通过过滤处理的配对样本进行了比较。两组UCB样本的过滤，平均体积分别为87.8和120.7 ml，有核细胞（NC）含量分别为9.7和23.8×10(8)，导致单核细胞（[MNCs] 74.2%-77.5%）、CD34(+)细胞（76.3%-79.0%）和集落形成细胞（64.1%-86.3%）有足够的平均细胞回收率。此外，我们检测到移植中红细胞（RBCs，89.2%-90.0%）和血小板（[PLTs] 77.5%-86.1%）有显著减少。相比之下，使用BC处理的平均减少率在PLTs（10%，p = 0.03）和RBCs（39.6%，p < 0.01）方面被证明有显著差异。与BC组中MNCs的较小显著增加（p < 0.05）相比，过滤后NC组成显示MNCs有高度显著增加，粒细胞减少（p < 0.01）。对于移植了WB来源祖细胞的小鼠，我们观察到骨髓中人类CD45(+)细胞的平均比例为15.3%±15.5%，而移植了过滤样本的小鼠为19.9%±16.8%（p = 0.03）。WB样本中人类CD34(+)细胞的平均比例为4.2%±3.1%，过滤样本为4.5%±3.2%（p = 0.68）。由于NOD/SCID小鼠移植数据表明过滤处理的HPC有显著的植入能力，未发现过滤的UCB细胞与WB移植中未减少体积的细胞相比对植入潜力有负面影响。StemQuick((TM))E过滤装置被证明是一种符合药品生产质量管理规范的从UCB中富集HPC和MNC的有用工具。过滤能够快速、标准化地制备体积减少的UCB移植，包括部分去除粒细胞、RBCs和PLTs，而无需离心。因此，经过过滤处理的UCB移植在人类中也很可能导致足够的造血重建。
Zotero 插件