Rim Yeri Alice, Nam Yoojun, Ju Ji Hyeon
CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea.
Division of Rheumatology, Department of Internal Medicine, Seoul St. Marys Hospital, Institute of Medical Science, College of Medicine, The Catholic University of Korea;
J Vis Exp. 2016 Dec 21(118):54650. doi: 10.3791/54650.
The recent development of human induced pluripotent stem cells (hiPSCs) proved that mature somatic cells can return to an undifferentiated, pluripotent state. Now, reprogramming is done with various types of adult somatic cells: keratinocytes, urine cells, fibroblasts, etc. Early experiments were usually done with dermal fibroblasts. However, this required an invasive surgical procedure to obtain fibroblasts from the patients. Therefore, suspension cells, such as blood and urine cells, were considered ideal for reprogramming because of the convenience of obtaining the primary cells. Here, we report an efficient protocol for iPSC generation from peripheral blood mononuclear cells (PBMCs). By plating the transduced PBMCs serially to a new, matrix-coated plate using centrifugation, this protocol can easily provide iPSC colonies. This method is also applicable to umbilical cord blood mononuclear cells (CBMCs). This study presents a simple and efficient protocol for the reprogramming of PBMCs and CBMCs.
人类诱导多能干细胞(hiPSC)的最新进展证明,成熟的体细胞可以恢复到未分化的多能状态。目前,重编程可通过多种类型的成人体细胞来完成:角质形成细胞、尿液细胞、成纤维细胞等。早期实验通常使用真皮成纤维细胞进行。然而,这需要通过侵入性手术从患者身上获取成纤维细胞。因此,诸如血液和尿液细胞等悬浮细胞由于获取原代细胞的便利性而被认为是重编程的理想选择。在此,我们报告了一种从外周血单个核细胞(PBMC)生成iPSC的高效方案。通过使用离心将转导后的PBMC依次接种到新的、基质包被的培养皿中,该方案能够轻松获得iPSC集落。此方法也适用于脐带血单个核细胞(CBMC)。本研究提出了一种用于PBMC和CBMC重编程的简单高效方案。