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新型体外分离系统在非细胞因子刺激供者中的应用:首次采集结果及后续冻存评估。

Leukapheresis in non-cytokine-stimulated donors with a new apheresis system: first-time collection results and evaluation of subsequent cryopreservation.

机构信息

Transfusion Medicine and Hemostaseology Department, University Hospital Erlangen, Erlangen, Germany.

出版信息

Transfusion. 2013 Apr;53(4):747-56. doi: 10.1111/j.1537-2995.2012.03787.x. Epub 2012 Jul 15.

Abstract

BACKGROUND

Adoptive cell therapy based on mononuclear cells (MNCs) became an important modality of cancer immunotherapy. Data about collection results and donor response of leukapheresis with the Spectra Optia v.5.0 (Terumo BCT) in nonmobilized donors are required.

STUDY DESIGN AND METHODS

Twelve MNC collections were performed using the Spectra Optia v.5.0 in non-cytokine-stimulated donors. Leukapheresis products and peripheral blood samples from donors were assayed for CD45+, CD34+, CD3+, and CD14+ cells by flow cytometry. Prefreeze and postthaw cell counts, cell viability, and numbers of colony-forming units were assessed in cryobags and compared to data from cryovials.

RESULTS

Leukapheresis yielded a mean of 5.26×10(9) ±2.2×10(9) CD45+ cells, 1.5×10(9) ±0.77×10(9) CD14+ monocytes, and 2.28×10(9) ±1.2×10(9) CD3+ Tcells by processing 6690±930mL of whole blood. A significant positive correlation between yield of CD3+ Tcells and residual platelets (PLTs) and red blood cells (RBCs) was observed. This did not apply for CD34+ and CD14+ white blood cell subsets. Mean collection efficiencies for CD14+ monocytes and CD3+ Tcells were 61.8±17 and 37.2±18%, respectively. Recovery of CD14+ cells after cryopreservation was 75.2±8.2%, which was significantly lower than recovery of CD45+ cells (81.4±5.5%; p=0.01).

CONCLUSION

This study of a small cohort demonstrates that the Spectra Optia v.5.0 is capable of collecting low product volumes with satisfactory MNC yields and low residual RBCs and PLTs in non-cytokine-mobilized apheresis. Our data suggest that cryovials can serve as a representative surrogate for the primary product cryobag.

摘要

背景

基于单核细胞(MNC)的过继细胞疗法已成为癌症免疫疗法的重要手段。需要有关 Spectra Optia v.5.0(Terumo BCT)在非动员供体中采集结果和供体反应的数据。

研究设计与方法

对 12 例非细胞因子刺激供体进行 Spectra Optia v.5.0 采集。通过流式细胞术检测供体的白细胞分离产物和外周血样本中的 CD45+、CD34+、CD3+和 CD14+细胞。在冷冻袋中评估预冷冻和冻融后的细胞计数、细胞活力和集落形成单位数量,并与冷冻管中的数据进行比较。

结果

通过处理 6690±930mL 全血,白细胞分离术获得了 5.26×10(9)±2.2×10(9)个 CD45+细胞、1.5×10(9)±0.77×10(9)个 CD14+单核细胞和 2.28×10(9)±1.2×10(9)个 CD3+T 细胞。观察到 CD3+T 细胞产量与残留血小板(PLT)和红细胞(RBC)之间存在显著正相关。这不适用于 CD34+和 CD14+白细胞亚群。CD14+单核细胞和 CD3+T 细胞的平均采集效率分别为 61.8±17%和 37.2±18%。冷冻保存后 CD14+细胞的回收率为 75.2±8.2%,明显低于 CD45+细胞的回收率(81.4±5.5%;p=0.01)。

结论

这项对小队列的研究表明,Spectra Optia v.5.0 能够在非细胞因子动员的白细胞分离中采集低体积产品,获得令人满意的 MNC 产量,并使 RBC 和 PLT 残留量低。我们的数据表明,冷冻管可以作为原始产品冷冻袋的代表性替代物。

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