大量溶解程序改变了目标细胞群体的数量.
Laura G Rico1, Roser Salvia1, Michael D Ward2
1Functional Cytomics Lab, Germans Trias i Pujol Research Institute (IGTP), ICO-Hospital Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona (Barcelona), Spain.
概括
大量溶解 (BL) 试验用于高灵敏度细胞分析,如可测量的残留疾病 (MRD) 检测,导致显著的细胞损失. 最小样本扰动 (MSP) 协议保留了更多的细胞,提高了流细胞计中罕见细胞检测的准确性.
科学领域:
- 血液学 血液学 血液学
- 免疫学 免疫学 免疫学
- 细胞生物学 细胞生物学
背景情况:
- 通过流细胞计 (FCM) 高灵敏细胞测量 (<10^5个细胞中的1个) 需要分析数百万个细胞,特别是用于可测量的残留疾病 (MRD) 检测.
- 传统的免疫类型定型协议和批量溶解 (BL) 试验往往无法获得足够的细胞数量,并可能导致显著的细胞损失.
- 建议采用最小样本干扰 (MSP) 协议,以更好地保护细胞完整性,并避免样本准备期间的细胞损失.
研究的目的:
- 评估和比较批量溶解 (BL) 协议与最小样本干扰 (MSP) 协议的有效性,用于高灵敏度细胞分析.
- 评估BL协议对细胞计数的影响,特别是对于MRD检测中的罕见细胞群.
主要方法:
- 通过将周围血液中 EGFP 表达K562细胞的已知百分比增加,创建了一个MRD模型.
- 样品使用批量溶解 (BL) 和最小样品扰动 (MSP) 协议进行处理.
- 流细胞计 (FCM) 用于分析和比较两个协议之间的细胞群.
主要成果:
- 在所有测试百分比中,BL样本与MSP样本相比,K562细胞种群的显著减少被观察到.
- 大量溶解 (BL) 协议导致与MSP协议相比,非死细胞的显著损失.
- 这些发现表明,在使用BL方法时,可能会高估或低估罕见的细胞.
结论:
- 大量溶解 (BL) 协议显著减少细胞数量,影响高灵敏度测量的准确性,特别是在MRD检测中的罕见细胞.
- 与BL方法相比,最小样本扰动 (MSP) 协议显示出优异的细胞恢复和保存.
- 批量溶解试剂和溶液的改进是必要的,以提高细胞恢复,并确保高灵敏度FCM应用的准确量化.
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