Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, University of Cambridge, Cambridge CB2 0QQ, UK.
Stem Cell Reports. 2015 Oct 13;5(4):660-71. doi: 10.1016/j.stemcr.2015.08.012. Epub 2015 Sep 17.
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.
人原代组织样本的采集地点和接收实验室通常不在同一地点，在人诱导多能干细胞（hiPSCs）衍生的地方。因此，在衍生之前样本的稳定性限制了采集地点和接收实验室之间的距离。为了研究样本的稳定性，我们采集了血液，并在室温下放置 5、24 或 48 小时，然后分离外周血单核细胞（PBMCs）并将其重编程为 iPSCs。此外，在 5 小时和 48 小时时间点的 PBMC 样本在液氮中冷冻 4 个月并被重编程为 iPSCs。所有时间点衍生的 hiPSC 系均具有多能性，染色体异常率无明显差异，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 小时低 3 倍和 10 倍；PBMCs 的冻融过程对重编程效率没有明显影响。
