人类骨髓 stromal 细胞:抗凝剂对干细胞特性的影响
Michaela Ferencakova1, Andrea Benova1,2, Ivan Raska3
1Laboratory of Molecular Physiology of Bone, Institute of Physiology of the Czech Academy of Sciences, Prague, Czechia.
Frontiers in cell and developmental biology
|October 4, 2023
概括
像EDTA或肝素这样的抗凝剂不会显著改变骨髓 stromal 细胞 (BMSCs). 然而,体外培养实质性地改变了BMSC基因表达,影响了它们在再生医学中的使用.
科学领域:
- 干细胞生物学 干细胞生物学
- 再生医学是一种再生医学.
- 分子生物学分子生物学
背景情况:
- 骨髓 stromal 细胞 (BMSCs) 是再生医学的关键多能干细胞.
- 目前的隔离方法使用抗凝剂,可能会改变BMSC的特征.
- 对于临床应用来说,研究BMSC分子谱的抗凝剂作用至关重要.
研究的目的:
- 确定乙烯基二胺三酸 (EDTA) 与肝素抗凝剂对人类BMSC隔离和特征的影响.
- 为了评估由于抗凝剂和随后的体外培养而导致的BMSCs的转录组变化.
主要方法:
- 从捐赠者 (48-85岁) 收集的人类骨髓吸附物使用EDTA或氨酸涂层管.
- 通过流细胞计,RNA-seq,CFU测定,增殖,分化,生物能学,代谢学,免疫染和RT-qPCR分析了BMSC.
- 来自同一患者的双对样本用于比较分析.
主要成果:
- 与EDTA相比,氨酸产生了更高的细胞计数和殖民地形成单位 (CFU).
- 在EDTA和肝素分离的BMSC之间没有观察到显著的分子特征差异.
- 在试验室培养显著改变了BMSC转录组,增加了核酸/细胞代谢基因,减少了炎症/染色质重塑基因,而不管抗凝剂.
结论:
- 抗凝剂的选择 (EDTA与肝素) 对BMSC分子特征和细胞组成的影响很小.
- 试管培养诱导了BMSCs的实质性转录基因转移,这是其临床和再生医学应用的关键考虑因素.
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