Vaskova Evgeniya A, Medvedev Sergey P, Sorokina Anastasiya E, Nemudryy Artem A, Elisaphenko Evgeniy A, Zakharova Irina S, Shevchenko Alexander I, Kizilova Elena A, Zhelezova Antonina I, Evshin Ivan S, Sharipov Ruslan N, Minina Julia M, Zhdanova Natalia S, Khegay Igor I, Kolpakov Fedor A, Sukhikh Gennadiy T, Pokushalov Evgeniy A, Karaskov Alexander M, Vlasov Valentin V, Ivanova Ludmila N, Zakian Suren M
1 The Federal Research Center Institute of Cytology and Genetics , the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia .
2 State Research Institute of Circulation Pathology, Ministry of Healthcare of the Russian Federation, Novosibirsk, Russia .
Stem Cells Dev. 2015 Dec 15;24(24):2912-24. doi: 10.1089/scd.2015.0204. Epub 2015 Sep 29.
Rat pluripotent stem cells, embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs) as mouse and human ones have a great potential for studying mammalian early development, disease modeling, and evaluation of regenerative medicine approaches. However, data on pluripotency realization and self-renewal maintenance in rat cells are still very limited, and differentiation protocols of rat ESCs (rESCs) and iPSCs to study development and obtain specific cell types for biomedical applications are poorly developed. In this study, the RNA-Seq technique was first used for detailed transcriptome characterization in rat pluripotent cells. The rESC and iPSC transcriptomes demonstrated a high similarity and were significantly different from those in differentiated cells. Additionally, we have shown that reprogramming of rat somatic cells to a pluripotent state was accompanied by X-chromosome reactivation. There were two active X chromosomes in XX rESCs and iPSCs, which is one of the key attributes of the pluripotent state. Differentiation of both rESCs and iPSCs led to X-chromosome inactivation (XCI). The dynamics of XCI in differentiating rat cells was very similar to that in mice. Two types of facultative heterochromatin described in various mammalian species were revealed on the rat inactive X chromosome. To explore XCI dynamics, we established a new monolayer differentiation protocol for rESCs and iPSCs that may be applied to study different biological processes and optimized for directed derivation of specific cell types.
大鼠多能干细胞、胚胎干细胞（ESCs）以及诱导多能干细胞（iPSCs），如同小鼠和人类的这些细胞一样，在研究哺乳动物早期发育、疾病建模以及评估再生医学方法方面具有巨大潜力。然而，关于大鼠细胞中多能性实现和自我更新维持的数据仍然非常有限，并且用于研究发育以及获得用于生物医学应用的特定细胞类型的大鼠胚胎干细胞（rESCs）和诱导多能干细胞的分化方案也很不完善。在本研究中，RNA测序技术首次被用于大鼠多能细胞的详细转录组特征分析。大鼠胚胎干细胞和诱导多能干细胞的转录组显示出高度相似性，并且与分化细胞中的转录组显著不同。此外，我们已经表明，大鼠体细胞重编程为多能状态伴随着X染色体重新激活。在XX大鼠胚胎干细胞和诱导多能干细胞中有两条活跃的X染色体，这是多能状态的关键特征之一。大鼠胚胎干细胞和诱导多能干细胞的分化都导致了X染色体失活（XCI）。分化中的大鼠细胞中XCI的动态变化与小鼠中的非常相似。在大鼠失活的X染色体上发现了在各种哺乳动物物种中描述的两种兼性异染色质。为了探索XCI动态变化，我们为大鼠胚胎干细胞和诱导多能干细胞建立了一种新的单层分化方案，该方案可用于研究不同的生物学过程，并针对特定细胞类型的定向诱导进行了优化。
