Cryopreservation of siRNA-Treated Cells Is Feasible.

IF 4.7 2区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Melanie Sauer, Xavier Segarra-Visent, Leon Breuer, Vasileios Tzirtziganis, Tatyana Ryaykenen, David A Cooper, Dimas Echeverria, Anastasia Kremer, Reka A Haraszti
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引用次数: 0

Abstract

Cryopreservation is a routine step in the manufacturing process of adoptive cell therapies (ACT), providing critical logistic flexibility. RNA interference (RNAi)-based therapies are increasingly being explored as enhancers or modulators of ACT. However, the impact of cryopreservation on cells treated with RNAi-based therapies has not been investigated before. In this study, we addressed this knowledge gap by examining silencing efficacy in small interfering RNA (siRNA)-treated cells that undergo cryopreservation. Our findings demonstrate that silencing in cryopreserved cells is comparable to that in cells maintained continuously in culture. Moreover, we found that the duration of siRNA exposure plays a significant role in cells that later undergo cryopreservation, with extended exposure improving silencing efficiency. However, this effect diminishes at higher siRNA concentrations. Additionally, we showed that siRNA treatment is feasible at low temperatures (2°C-8°C), and siRNA-treated cells can be cryopreserved for extended periods (at least 1 month) without loss of efficacy. Our work establishes the feasibility of integrating siRNA treatments into current manufacturing processes for ACT.

sirna处理细胞的低温保存是可行的。
低温保存是过继细胞疗法(ACT)生产过程中的常规步骤,提供了关键的物流灵活性。基于RNA干扰(RNAi)的治疗方法越来越多地被探索为ACT的增强剂或调节剂。然而,冷冻保存对以rnai为基础的治疗方法处理的细胞的影响之前尚未研究过。在这项研究中,我们通过检测小干扰RNA (siRNA)处理的细胞在低温保存中的沉默效果来解决这一知识差距。我们的研究结果表明,冷冻保存细胞的沉默与连续培养细胞的沉默相当。此外,我们发现siRNA暴露的持续时间在随后进行冷冻保存的细胞中起着重要作用,延长暴露时间可以提高沉默效率。然而,这种效应在siRNA浓度较高时减弱。此外,我们发现siRNA处理在低温(2°C-8°C)下是可行的,并且siRNA处理的细胞可以长时间(至少1个月)冷冻保存而不会失去功效。我们的工作确定了将siRNA处理整合到当前ACT制造工艺中的可行性。
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
Nucleic acid therapeutics
Nucleic acid therapeutics BIOCHEMISTRY & MOLECULAR BIOLOGY-CHEMISTRY, MEDICINAL
CiteScore
7.60
自引率
7.50%
发文量
47
审稿时长
>12 weeks
期刊介绍: Nucleic Acid Therapeutics is the leading journal in its field focusing on cutting-edge basic research, therapeutic applications, and drug development using nucleic acids or related compounds to alter gene expression. The Journal examines many new approaches for using nucleic acids as therapeutic agents or in modifying nucleic acids for therapeutic purposes including: oligonucleotides, gene modification, aptamers, RNA nanoparticles, and ribozymes.
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