中国输血杂志2025，Vol.38Issue(5)：723-733,11.DOI:10.13303/j.cjbt.issn.1004-549x.2025.05.020
从滤白残留物中回收单个核细胞的稳定性研究
Study on the stability of PBMCs recovered from leukocyte-depleted filter residues
摘要
Abstract
Objective To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell(PBMC)recovery with high viability,long-term storage stability,and sterility of the harvested resi-dues,thereby providing a clinically translatable strategy.Methods Three sterile bag-packaged solutions—Saline,Solvent,and Hanks' balanced salt solution(HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters.Nucleat-ed cell recovery rate and viability of the harvested residues were compared.The optimal solution identified was applied to an expanded sample set.PBMC viability and yield were evaluated after 1h vs 48h storage of the residues.PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month,followed by post-thaw comparisons of viability and T-cell ex-pansion capacity.Results The Solvent group achieved the highest and most consistent nucleated cell recovery rate.Post-flush recovery rate from filters after 400 mL whole blood processing was(21.3±1.6)%for the Solvent group,significantly higher than Saline group(19.2±6.3)%and HBSS group(11.2±5.0)%,with residues from all groups maintaining viabili-ty>90%.No biologically significant difference in residue viability was observed between 48h vs 1h storage groups(93.3±2.3)%vs(95.7±1.8)%).PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups[(48.2%±9.5%)vs(40.41%±8.35%),P>0.05],with(17.7±2.6)×107 cells.After 1-month cryopreservation and 10-day expansion,PBMCs isolated from 48-hour-stored residues retained(91.2±3.2)%viability and achieved a(61.9±15.9)-fold expansion.Conclusion The bag-packaged Solvent,as a back-flush solution,enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections.These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃,with post-cryopreservation(1-month liquid nitrogen)viability and expansion ca-pacity remaining stable.This protocol complies with blood bank regulatory criteria,addresses the concerns about the infec-tious window period in cell therapy raw materials,and provides a clinically translatable strategy for PBMC-based applica-tions.关键词
白细胞滤器/反冲溶液/滤白残留物/外周血单个核细胞Key words
leukocyte filter/backflush solution/leukocyte-depleted filter residues/peripheral blood mononuclear cells(PBMCs)分类
医药卫生引用本文复制引用
林菊,相志强,杜东芬,袁芳,王妙玉,吴越,黄凯玉,桑列勇..从滤白残留物中回收单个核细胞的稳定性研究[J].中国输血杂志,2025,38(5):723-733,11.基金项目
绍兴市卫生健康科技计划项目(2023SKY096) （2023SKY096）
