Assessment of genetic and epigenetic variation in human IPS cells-RNA

In order to progress human induced pluripotent stem cells (hiPSCs) towards the clinic, several outstanding questions must be addressed. It is possible to reprogram different somatic cell types into hiPSCs and from studies in the mouse, it appears that an epigenetic memory of the starting cell type is carried over to hiPSCs. However a comprehensive comparative study of the characteristics of these hiPSCs has been missing from the literature. Importantly studies which aimed to address these aspects of hiPSCs have used cells from different patients. In order to avoid this important confounding variable and to keep the genetic background constant, tissue samples were procured from the patients and reprogrammed to iPS cells. The transcriptomes of these iPS cells will be compared.Protocol: primary cultures of cells were reprogrammed to iPS cells. RNA was extracted using a standard column extraction kit.

为了推进由人类诱导的多能干细胞（hiPSCs）向临床应用的进程，必须解决一系列关键问题。将不同类型的体细胞重编程为hiPSCs是可行的，而根据小鼠研究，起始细胞类型的表观遗传记忆似乎被传递到了hiPSCs中。然而，关于这些hiPSCs特性的全面比较研究在文献中尚属缺失。值得注意的是，旨在解决这些hiPSCs方面的研究使用了来自不同患者的细胞。为了避免这一重要的混杂变量并保持遗传背景的稳定性，从患者处采集了组织样本并重编程为iPS细胞。这些iPS细胞的转录组将被进行比较。实验方法：首先将细胞原代培养重编程为iPS细胞。使用标准柱式提取试剂盒提取RNA。