PO.MCB07.04 · 分子与细胞生物学

RNA-seq分析证实ThawReady™ THP-1即用型检测细胞的解冻后转录组稳定性

RNA-seq analysis confirms post-thaw transcriptomic stability in ThawReady (TM) THP-1 assay-ready cells

海报缩略图:RNA-seq分析证实ThawReady™ THP-1即用型检测细胞的解冻后转录组稳定性
编号 7346 展板 3 时间 4/22 09:00–12:00 区域 Section 24 主讲 Ajeet Singh, PhD
分会场 Transcription Factor Function in Cell Identity, Signaling, and Post-Transcriptional Control
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作者与单位 Authors & Affiliations

Ajeet P. Singh, Utsav Sharma, Lucas Underwood, Noah Wax, Steve King, Nilay Chakraborty, Jonathan Jacobs

ATCC, Manassas, VA

摘要 Abstract

中文摘要
冷冻保存的细胞系已成为高通量筛选和检测开发的必备工具,可在各实验之间提供便利性和一致性。然而,冻融过程可能诱导细胞应激,从而潜在地影响基因表达并损害功能可靠性。为确保检测性能的一致性,验证冷冻保存的细胞在解冻后是否保留其转录组和功能完整性至关重要。在本研究中,我们使用RNA测序(RNA-seq)评估了ThawReady™ THP-1(TIB-202-AR™)细胞的转录组稳定性。在解冻后立即(0小时)以及2小时和8小时恢复间隔后对细胞进行分析,并与新鲜培养的THP-1细胞进行比较。主成分分析和层次聚类分析显示,解冻后样本与新鲜对照紧密聚集,表明细胞身份和整体基因表达模式得到保留。基因表达谱在THP-1细胞特有的关键免疫和炎症通路上表现出高度一致性。差异表达分析以绝对倍数变化>5和FDR校正p值<0.05为阈值,鉴定出解冻后相对于新鲜培养在0小时改变的基因178个、2小时951个、8小时713个。尽管存在这些短暂变化,与细胞存活和增殖相关的通路仍保持稳定。值得注意的是,吞噬体形成是富集程度最高的通路,提示存在支持细胞活力的适应性恢复反应。总体而言,这些结果证实ThawReady™ THP-1细胞在解冻后维持稳健的转录组完整性,支持其在下游免疫和炎症相关功能检测中的可靠性和可重复性。
查看英文原文 English abstract
Cryopreserved cell lines have become essential for high-throughput screening and assay development, offering convenience and consistency across experiments. However, the freeze-thaw process can induce cellular stress, potentially affecting gene expression and compromising functional reliability. To ensure consistent assay performance, it is crucial to verify that cryopreserved cells retain their transcriptomic and functional integrity after thawing. In this study, we evaluated the transcriptomic stability of ThawReady™ THP-1 (TIB-202-AR™) cells using RNA sequencing (RNA-seq). Cells were analyzed immediately after thawing (0-hour) and following 2-hour and 8-hour recovery intervals and were compared to freshly cultured THP-1 cells. Principal component and hierarchical clustering analyses revealed that post-thaw samples closely grouped with fresh controls, demonstrating preservation of cellular identity and global gene expression patterns. Gene expression profiles showed strong concordance across key immune and inflammatory pathways characteristic of THP-1 cells. Differential expression analysis identified 178 genes altered at 0 hours, 951 at 2 hours, and 713 at 8 hours post-thaw relative to fresh culture, using a threshold of absolute fold change >5 and FDR-adjusted p-value <0.05. Despite these transient changes, pathways related to cell survival and proliferation remained stable. Notably, phagosome formation was the top-enriched pathway, suggesting adaptive recovery responses that support cell viability. Overall, these results confirm that ThawReady™ THP-1 cells maintain robust transcriptomic integrity post-thaw, supporting their reliability and reproducibility for downstream immune and inflammation-related functional assays.
利益披露 Disclosure
A. P. Singh, None.. U. Sharma, None.. L. Underwood, None.. N. Wax, None.. S. King, None.. N. Chakraborty, None.. J. Jacobs, None.

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