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测试HPC和MNC的稳定性
Jinxia Ma1, Lipei Shao1, Tatyana Fuksenko1
1Center for Cellular Engineering, Department of Transfusion Medicine and Center for Cellular Engineering, NIH Clinical Center, Bethesda, MD 20892, USA.
血造细胞和单核细胞 (MNC) 样本在4°C下稳定3天,用于WBC计数和流细胞计量. 然而,HPC显示了早期的转录基因变化,需要立即进行分子分析.
科学领域:
- 细胞和分子生物学
- 血液学
- 生物技术
背景情况:
- 血造原生细胞 (HPC) 和单核细胞 (MNC) 对于骨髓移植等细胞疗法至关重要.
- 在采集,储存和运输过程中评估产品特性对于维持细胞活力和功能至关重要.
研究的目的:
- 评估在4°C保存长达4天的新鲜HPC和MNC产品样本的功能和分子稳定性
- 确定存储时间对细胞活力,表型标志物和转录组特征的影响.
主要方法:
- 在4°C下保存HPC (n=7) 和MNC (n=6) 的样本,并在0~4天进行分析.
- 评估包括白细胞计数,流细胞计量 (使用特定的细胞表面标记物) 和大量RNA测序 (RNA-seq).
- 通过使用7-AAD染色来评估细胞活力.
主要成果:
- 白细胞计数和流细胞计量标记在长达4天内保持相对稳定,HPC的变异性更大.
- 细胞活力随着时间的推移显著下降,显示出产品间的变异性.
- 在MNC样本中保持稳定的基因表达特征,而HPC样本在存储的第一天显示出显著的转录组变化.
结论:
- 在4°C下,白细胞计数和流细胞计量标记在长达3天内保持稳定,支持延迟的表型分析.
- 在HPC中显著的转录组变化需要立即进行分子评估以保持完整性.
- 尽管表型标记稳定,但研究结果强调了基于HPC的应用需要及时进行分子测试.
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