O'Brien Carmel M, Chy Hun S, Zhou Qi, Blumenfeld Shiri, Lambshead Jack W, Liu Xiaodong, Kie Joshua, Capaldo Bianca D, Chung Tung-Liang, Adams Timothy E, Phan Tram, Bentley John D, McKinstry William J, Oliva Karen, McMurrick Paul J, Wang Yu-Chieh, Rossello Fernando J, Lindeman Geoffrey J, Chen Di, Jarde Thierry, Clark Amander T, Abud Helen E, Visvader Jane E, Nefzger Christian M, Polo Jose M, Loring Jeanne F, Laslett Andrew L
Clayton and Parkville, CSIRO Manufacturing, Victoria, Australia.
Australian Regenerative Medicine Institute, Monash University, Clayton, Victoria, Australia.
Stem Cells. 2017 Mar;35(3):626-640. doi: 10.1002/stem.2558. Epub 2017 Jan 19.
The study and application of human pluripotent stem cells (hPSCs) will be enhanced by the availability of well-characterized monoclonal antibodies (mAbs) detecting cell-surface epitopes. Here, we report generation of seven new mAbs that detect cell surface proteins present on live and fixed human ES cells (hESCs) and human iPS cells (hiPSCs), confirming our previous prediction that these proteins were present on the cell surface of hPSCs. The mAbs all show a high correlation with POU5F1 (OCT4) expression and other hPSC surface markers (TRA-160 and SSEA-4) in hPSC cultures and detect rare OCT4 positive cells in differentiated cell cultures. These mAbs are immunoreactive to cell surface protein epitopes on both primed and naive state hPSCs, providing useful research tools to investigate the cellular mechanisms underlying human pluripotency and states of cellular reprogramming. In addition, we report that subsets of the seven new mAbs are also immunoreactive to human bone marrow-derived mesenchymal stem cells (MSCs), normal human breast subsets and both normal and tumorigenic colorectal cell populations. The mAbs reported here should accelerate the investigation of the nature of pluripotency, and enable development of robust cell separation and tracing technologies to enrich or deplete for hPSCs and other human stem and somatic cell types. Stem Cells 2017;35:626-640.
能够检测细胞表面表位的特性明确的单克隆抗体(mAb)的可得性,将促进人类多能干细胞(hPSC)的研究与应用。在此,我们报告了七种新单克隆抗体的产生,这些抗体可检测活的和固定的人类胚胎干细胞(hESC)及人类诱导多能干细胞(hiPSC)上存在的细胞表面蛋白,证实了我们之前的预测,即这些蛋白存在于hPSC的细胞表面。这些单克隆抗体在hPSC培养物中均与POU5F1(OCT4)表达及其他hPSC表面标志物(TRA-160和SSEA-4)高度相关,并能检测分化细胞培养物中罕见的OCT4阳性细胞。这些单克隆抗体对始发态和原始态hPSC的细胞表面蛋白表位均具有免疫反应性,为研究人类多能性及细胞重编程状态的细胞机制提供了有用的研究工具。此外,我们报告这七种新单克隆抗体的亚群对人类骨髓间充质干细胞(MSC)、正常人乳腺亚群以及正常和致瘤性结肠直肠细胞群体也具有免疫反应性。本文报道的单克隆抗体应能加速对多能性本质的研究,并推动强大的细胞分离和追踪技术的发展,以富集或去除hPSC及其他人类干细胞和体细胞类型。《干细胞》2017年;35卷:626 - 640页