Quality Assessment of Cryopreserved Peripheral Blood Stem Cell Products: Evaluation of Two Methods for Flow Cytometric Viability Testing

IF 2.2 4区医学 Q3 HEMATOLOGY International Journal of Laboratory Hematology Pub Date : 2024-09-22 DOI:10.1111/ijlh.14374

Vladimira Rimac, Ines Bojanić, Marijana Škifić, Sanja Dabelić, Branka Golubić Ćepulić

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Abstract

Introduction

The standard flow cytometry method for viability testing using 7-aminoactinomycin D (7-AAD) determines cells in necrosis and late apoptosis. The colony-forming unit (CFU) assay, which evaluates the proliferation ability of HSCs, is also used in graft quality assessment despite known deficiencies that make this assay impractical in routine clinical settings. The aim was to compare the effectiveness of the flow cytometry 7-AAD/annexin V method with the 7-AAD method in assessing the quality of HSCs in autologous and allogeneic peripheral blood stem cell (PBSC) products.

Methods

Thirty autologous and 30 allogeneic fresh and thawed cryopreserved PBSC products were included in this study. The viability of HSCs was determined using the 7-AAD method and 7-AAD/annexin V method on a flow cytometer, while their clonogenic capacity was assessed by CFU assay.

Results

There was an excellent correlation for CD34+ cell viability between the 7-AAD and the 7-AAD/annexin V method for fresh samples (Rs = 0.930, p < 0.001) and a good correlation for thawed PBSC samples (Rs = 0.739, p < 0.001). Excellent correlation was observed for post-thaw CD34+ cell recovery between the two methods for viability (Rs = 0.980, p < 0.001). Statistical analysis showed a weak correlation between CFU-GM recovery and CD34+ cell recovery, regardless of which viability testing method was used (7-AAD method p = 0.021, Rs = 0.298; 7-AAD/annexin V method p = 0.029, Rs = 0.282).

Conclusions

Results of this study showed that in the quality assessment of cryopreserved PBSC product viability, the 7-AAD/annexin V method had no added value compared to the 7-AAD method, which was suitable enough for routine quality control of cryopreserved autologous and allogeneic PBSC samples.

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冷冻保存的外周血干细胞产品的质量评估：两种流式细胞活力测试方法的评估。

简介使用 7-aminoactinomycin D（7-AAD）进行活力检测的标准流式细胞术方法可确定细胞坏死和晚期凋亡。集落形成单位（CFU）检测法可评估造血干细胞的增殖能力，也被用于移植物质量评估，尽管这种检测法存在已知的缺陷，在常规临床环境中并不实用。我们的目的是比较流式细胞术 7-AAD/annexin V 法与 7-AAD 法在评估自体和异体外周血干细胞（PBSC）产品中造血干细胞质量方面的有效性：方法：本研究纳入了 30 个自体和 30 个异体新鲜和解冻冷冻保存的 PBSC 产品。在流式细胞仪上用 7-AAD 法和 7-AAD/annexin V 法测定造血干细胞的存活率，用 CFU 法评估造血干细胞的克隆能力：结果：对于新鲜样本，7-AAD 法和 7-AAD/annexin V 法对 CD34+ 细胞存活率有很好的相关性（Rs = 0.930，p 结论：7-AAD 法和 7-AAD/annexin V 法对 CD34+ 细胞存活率有很好的相关性（Rs = 0.930，p 结论）：本研究结果表明，在冷冻保存的 PBSC 产品存活率的质量评估中，7-AAD/附件素 V 法与 7-AAD 法相比没有附加值，7-AAD 法适用于冷冻保存的自体和异体 PBSC 样品的常规质量控制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Laboratory Hematology 医学-血液学

CiteScore

4.50

自引率

6.70%

发文量

211

审稿时长

6-12 weeks

期刊介绍： The International Journal of Laboratory Hematology provides a forum for the communication of new developments, research topics and the practice of laboratory haematology. The journal publishes invited reviews, full length original articles, and correspondence. The International Journal of Laboratory Hematology is the official journal of the International Society for Laboratory Hematology, which addresses the following sub-disciplines: cellular analysis, flow cytometry, haemostasis and thrombosis, molecular diagnostics, haematology informatics, haemoglobinopathies, point of care testing, standards and guidelines. The journal was launched in 2006 as the successor to Clinical and Laboratory Hematology, which was first published in 1979. An active and positive editorial policy ensures that work of a high scientific standard is reported, in order to bridge the gap between practical and academic aspects of laboratory haematology.

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