Yamaguchi Tomoyuki, Hamanaka Sanae, Nakauchi Hiromitsu
Centre for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan,
Methods Mol Biol. 2014;1210:143-50. doi: 10.1007/978-1-4939-1435-7_11.
This chapter describes a newly developed method for generating and maintaining rat induced pluripotent stem cells (riPSCs). We first provide a detailed protocol for the generation of lentiviral vector carrying three reprogramming factors to produce high-quality riPSCs. This technique allows reprogramming of rat somatic cells to ground state with germ-line competence. Subsequently, we elaborate a detailed protocol for the generation of riPSCs from rat embryonic fibroblast (REF). Finally, the protocols for the optimal culture conditions of riPSCs and preparation of frozen stock are described. We also outline the advantages of generating riPSCs.
本章介绍了一种新开发的用于生成和维持大鼠诱导多能干细胞（riPSC）的方法。我们首先提供了一个详细的方案，用于生成携带三种重编程因子的慢病毒载体，以产生高质量的riPSC。该技术可将大鼠体细胞重编程至具有种系能力的基态。随后，我们详细阐述了从大鼠胚胎成纤维细胞（REF）生成riPSC的方案。最后，描述了riPSC的最佳培养条件和冷冻保存液制备的方案。我们还概述了生成riPSC的优势。
