Schwandt Svenja, Korschgen Lutz, Peters Svenja, Kogler Gesine
Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Duesseldorf, Germany.
Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Duesseldorf, Germany.
Cytotherapy. 2016 May;18(5):642-52. doi: 10.1016/j.jcyt.2016.02.003.
Collection and processing characteristics influencing quality of cord blood (CB) units play an essential role to cord blood banks (CBBs). At many CBBs, volume reduction is performed using hydroxyethyl starch (HES) and the Sepax (Biosafe) automated cell processing system. Due to the withdrawal of HES from the European market, a validation of the nonHES protocol was performed.
This partially retrospective study identified CB characteristics such as gestational age and CB volume/cell count correlated with higher quality. For the nonHES validation, CB was analyzed for total nucleated cell (TNC), mononuclear cell (MNC) recovery, hematocrit (HCT) and colony-forming units (CFUs). Viabilities of CD34(+) and CD45(+) cells were determined by 7-aminoactinomycin D (7-AAD) and AnnexinV (AnnV) staining and compared for 21 mL and 42 mL buffy coat (BC) samples applying the HES/nonHES protocol.
Factors affecting the potency of CB transplants were the gestational age and the volume reduction to a defined BC volume. High initial cell counts and CB volumes correlated negatively with post-processing TNC recovery for lower BC volumes. Post-processing HES and nonHES results were comparable, but nonHES revealed a significantly lower post-thaw recovery of viable CD34(+) cells measured by 7-AAD/AnnV (21 mL: 45.4 ± 16.4%; 42 mL: 67.3 ± 14.5%) as compared with HES (21 mL: 72.7 ± 14.4%, P = 0.0164; 42 mL: 83.4 ± 14.7%, P = 0.0203).
Due to the lower post-thaw CD34(+) cell viability (AnnV(-)/7-AAD(-)) for nonHES samples, the use of HES is recommended, ideally combined with a high BC volume. The post-processing HCT has no statistically significant impact on the post-thaw CD34(+) cell viability (AnnV(-)/7-AAD(-)).
影响脐血（CB）单位质量的采集和处理特性对脐血库（CBB）至关重要。在许多脐血库中，使用羟乙基淀粉（HES）和Sepax（Biosafe）自动化细胞处理系统进行体积减少。由于HES退出欧洲市场，因此对非HES方案进行了验证。
这项部分回顾性研究确定了与较高质量相关的CB特征，如胎龄和CB体积/细胞计数。对于非HES验证，分析了CB的总核细胞（TNC）、单核细胞（MNC）回收率、血细胞比容（HCT）和集落形成单位（CFU）。通过7-氨基放线菌素D（7-AAD）和膜联蛋白V（AnnV）染色测定CD34(+)和CD45(+)细胞的活力，并比较应用HES/非HES方案的21 mL和42 mL血沉棕黄层（BC）样本。
影响CB移植效力的因素是胎龄和将体积减少至确定的BC体积。对于较低的BC体积，高初始细胞计数和CB体积与处理后TNC回收率呈负相关。处理后HES和非HES的结果具有可比性，但与HES相比，非HES显示通过7-AAD/AnnV测量的解冻后存活CD34(+)细胞的回收率显著降低（21 mL：45.4±16.4%；42 mL：67.3±14.5%）（HES：21 mL：72.7±14. %，P = 0.0164；42 mL：83.4±14.7%，P = 0.0203）。
由于非HES样本解冻后CD34(+)细胞活力（AnnV(-)/7-AAD(-)）较低，建议使用HES，理想情况下与高BC体积相结合。处理后HCT对解冻后CD34(+)细胞活力（AnnV(-)/7-AAD(-)）没有统计学上的显著影响。
