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Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr.
Stem Cell Reports ( IF 5.9 ) Pub Date : 2015-09-17 , DOI: 10.1016/j.stemcr.2015.08.012
Chukwuma A Agu 1 , Filipa A C Soares 2 , Alex Alderton 1 , Minal Patel 1 , Rizwan Ansari 1 , Sharad Patel 1 , Sally Forrest 1 , Fengtang Yang 1 , Jonathan Lineham 1 , Ludovic Vallier 3 , Christopher M Kirton 1
Affiliation  

The collection sites of human primary tissue samples and the receiving laboratories, where the human induced pluripotent stem cells (hIPSCs) are derived, are often not on the same site. Thus, the stability of samples prior to derivation constrains the distance between the collection site and the receiving laboratory. To investigate sample stability, we collected blood and held it at room temperature for 5, 24, or 48 hr before isolating peripheral blood mononuclear cells (PBMCs) and reprogramming into IPSCs. Additionally, PBMC samples at 5- and 48-hr time points were frozen in liquid nitrogen for 4 months and reprogrammed into IPSCs. hIPSC lines derived from all time points were pluripotent, displayed no marked difference in chromosomal aberration rates, and differentiated into three germ layers. Reprogramming efficiency at 24- and 48-hr time points was 3- and 10-fold lower, respectively, than at 5 hr; the freeze-thaw process of PBMCs resulted in no obvious change in reprogramming efficiency.



中文翻译:

从室温下保存长达 48 小时的血液样本成功生成人类诱导多能干细胞系。

人类原代组织样本的采集地点和人类诱导多能干细胞(hIPSC)的接收实验室通常不在同一地点。因此,样品在提取之前的稳定性限制了采集地点和接收实验室之间的距离。为了研究样品稳定性,我们收集血液并将其在室温下保存 5、24 或 48 小时,然后分离外周血单核细胞 (PBMC) 并重编程为 IPSC。此外,将 5 小时和 48 小时时间点的 PBMC 样品在液氮中冷冻 4 个月,并重新编程为 IPSC。来自所有时间点的 hIPSC 系都是多能的,染色体畸变率没有显着差异,并且分化为三个胚层。24 小时和 48 小时时间点的重编程效率分别比 5 小时时低 3 倍和 10 倍;PBMCs的冻融过程没有导致重编程效率的明显变化。

更新日期:2015-09-17
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