Jahan Suria, Maganti Harinad B, Patel Jaina, Pasha Roya, Doxtator Emily, Manesia Javed K, Pineault Nicolas
Canadian Blood Services, Innovation & Portfolio Management, Ottawa, Ontario, Canada.
Biochemistry, Microbiology and Immunology Department, University of Ottawa, Ottawa, Ontario, Canada.
Vox Sang. 2025 May;120(5):490-502. doi: 10.1111/vox.70012. Epub 2025 Mar 25.
Maximizing the potency of cord blood (CB) units (CBU) is a goal of all CB banks. Low transplant cell dose in CB transplantation is one of several contributors to the slower engraftment. Processing delays could also be at play because CBU can be stored at room temperature (RT) for up to 48 h before cryopreservation. We hypothesized that prolonged storage at RT reduces the potency and engraftment activities of CBU due to the loss of haematopoietic stem cells (HSCs) and progenitor cells.
CBU were split in half, with one half processed shortly after collection and the other half after 40 h of storage at RT. Viabilities and potencies were compared post thaw, and engraftment activity was tested in xenotransplants. Bioinformatic analysis was carried out to identify pathways modulated by prolonged storage at RT.
Prolonged storage at RT reduced the viability of CB cells and potency of grafts based on in vitro and in vivo assays. Transcriptomic analyses revealed that paracrine factors released in CBU induce several changes in gene expression programmes in CD34+ cells. These included the activation of the senescence gene CDKN1B and down-regulation of cell cycle and autophagy genes. Supplementation of CBU with trehalose prevented the loss in cell viability and potency. The latter was associated with the restoration of autophagy and suppression of the senescence gene CDKN1B.
These results stress the importance of rapid processing of CBU and support further investigations on trehalose as a natural supplement to protect CBU' potency during storage.
使脐血（CB）单位（CBU）的效力最大化是所有脐血库的目标。脐血移植中低移植细胞剂量是导致植入较慢的几个因素之一。处理延迟也可能起作用，因为CBU在冷冻保存前可在室温（RT）下储存长达48小时。我们假设，由于造血干细胞（HSC）和祖细胞的损失，在室温下长时间储存会降低CBU的效力和植入活性。
将CBU分成两半，一半在采集后不久进行处理，另一半在室温下储存40小时后处理。解冻后比较活力和效力，并在异种移植中测试植入活性。进行生物信息分析以确定室温下长时间储存所调节的途径。
基于体外和体内试验，在室温下长时间储存降低了CB细胞的活力和移植物的效力。转录组分析表明，CBU中释放的旁分泌因子在CD34+细胞中诱导了基因表达程序的几种变化。这些变化包括衰老基因CDKN1B的激活以及细胞周期和自噬基因的下调。用海藻糖补充CBU可防止细胞活力和效力的丧失。后者与自噬的恢复和衰老基因CDKN1B的抑制有关。
这些结果强调了快速处理CBU的重要性，并支持进一步研究海藻糖作为一种天然补充剂，以在储存期间保护CBU的效力。
