The Impact of Ammonium Chloride-Based Erythrocyte Lysis Process on Banked Adipose-Derived Stem Cells.

IF 1.4 4区 生物学 Q4 CELL BIOLOGY
Biopreservation and Biobanking Pub Date : 2022-06-01 Epub Date: 2021-10-26 DOI:10.1089/bio.2021.0035
Zifei Li, Dali Mu, Chunjun Liu, Minqiang Xin, Su Fu, Shangshan Li, Jun Qi, Hao Cheng, Chenglong Wang, Qian Wang, Jie Luan
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引用次数: 0

Abstract

The safety of banked human adipose-derived stem cells (hADSCs) purified by 155 mM ammonium chloride (NH4Cl)-based erythrocyte lysis has not been evaluated. This study was conducted to determine the impact of NH4Cl-based erythrocyte lysis on the biological characteristics of cryopreserved hADSCs. Stromal vascular fractions (SVFs) were obtained from lipoaspirates and purified with NH4Cl-based erythrocyte lysis (lysis group) or without (nonlysis group). The hADSCs were freshly isolated (fresh group) from SVFs and/or cryopreserved for 2 weeks (cryo group). The morphologies, immunophenotypes, viability, apoptosis, and growth kinetics of each group were compared. The cell cycle and differentiation capacity assays were performed in both cryopreserved groups. All groups showed similar cell morphology, immunological phenotypes, and viability. However, the main effect of lysis and its interaction with cryopreservation were observed when early apoptosis was regarded as a dependent variable in two-way repeated-measures analysis of variance. After cryopreservation, significant growth retardation and S-phase fraction reduction were observed in lytic hADSCs compared with those in nonlytic hADSCs. No significant differences in the adipogenic and osteogenic differentiation capacities were found between the two groups. Although NH4Cl-based erythrocyte lysis did not affect the cell morphology, immunological phenotypes, viability, and adipogenic and osteogenic differentiation capacities of cryopreserved hADSCs, exposure to NH4Cl-based erythrocyte lysis or its synergistic action with cryopreservation may induce apoptosis and inhibit the proliferation and mitosis of cryopreserved hADSCs. These results indicate that NH4Cl-based erythrocyte lysis is not suitable for high-quality banked collection of hADSCs for future clinical applications. Further development of safe, convenient, and cost-effective purification methods of hADSCs is warranted.

氯化铵基红细胞溶解过程对储存脂肪干细胞的影响。
以155 mM氯化铵(NH4Cl)为基础的红细胞溶解纯化的储存的人脂肪源性干细胞(hADSCs)的安全性尚未得到评估。本研究旨在确定nh4cl为基础的红细胞溶解对冷冻保存的hscs生物学特性的影响。从抽脂液中获得基质血管组分(svf),用nh4cl为基础的红细胞溶解(溶解组)或不溶解(非溶解组)纯化。从svf中新鲜分离的hscs(新鲜组)和/或冷冻保存2周(冷冻组)。比较各组细胞形态学、免疫表型、细胞活力、细胞凋亡和生长动力学。两组均进行细胞周期和分化能力测定。各组细胞形态、免疫表型和活力相似。然而,在双向重复测量方差分析中,当早期细胞凋亡被视为一个因变量时,观察到裂解的主要影响及其与低温保存的相互作用。冷冻保存后,与未裂解的hscs相比,裂解的hscs明显生长迟缓,s相分数减少。两组间的成脂和成骨分化能力无显著差异。虽然以nh4cl为基础的红细胞溶解不影响冷冻保存的hscs的细胞形态、免疫表型、活力以及成脂和成骨分化能力,但暴露于nh4cl为基础的红细胞溶解或其与冷冻保存的协同作用可能诱导细胞凋亡并抑制冷冻保存的hscs的增殖和有丝分裂。这些结果表明,基于nh4cl的红细胞溶解不适合用于未来临床应用的高质量hascs库收集。进一步开发安全、方便、经济高效的hscs纯化方法是必要的。
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来源期刊
Biopreservation and Biobanking
Biopreservation and Biobanking CELL BIOLOGY-MEDICAL LABORATORY TECHNOLOGY
CiteScore
3.30
自引率
12.50%
发文量
114
审稿时长
6-12 weeks
期刊介绍: Biopreservation and Biobanking is the first journal to provide a unifying forum for the peer-reviewed communication of recent advances in the emerging and evolving field of biospecimen procurement, processing, preservation and banking, distribution, and use. The Journal publishes a range of original articles focusing on current challenges and problems in biopreservation, and advances in methods to address these issues related to the processing of macromolecules, cells, and tissues for research. In a new section dedicated to Emerging Markets and Technologies, the Journal highlights the emergence of new markets and technologies that are either adopting or disrupting the biobank framework as they imprint on society. The solutions presented here are anticipated to help drive innovation within the biobank community. Biopreservation and Biobanking also explores the ethical, legal, and societal considerations surrounding biobanking and biorepository operation. Ideas and practical solutions relevant to improved quality, efficiency, and sustainability of repositories, and relating to their management, operation and oversight are discussed as well.
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