Czechanski Anne, Byers Candice, Greenstein Ian, Schrode Nadine, Donahue Leah Rae, Hadjantonakis Anna-Katerina, Reinholdt Laura G
Genetic Resource Science, The Jackson Laboratory, Bar Harbor, Maine, USA.
Developmental Biology Program, Sloan-Kettering Institute, New York, New York, USA.
Nat Protoc. 2014 Mar;9(3):559-74. doi: 10.1038/nprot.2014.030. Epub 2014 Feb 6.
Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbred mouse strains. Recently, there have been considerable advances in the field of embryonic stem cell biology, particularly in the area of pluripotency maintenance in the epiblast from which the mESCs are derived. Here we describe a protocol for efficient derivation of germline-competent mESCs from any mouse strain, including strains previously deemed nonpermissive. We provide a protocol that is generally applicable to most inbred strains, as well as a variant for nonpermissive strains. By using this protocol, mESCs can be derived in 3 weeks and fully characterized after an additional 12 weeks, at efficiencies as high as 90% and in any strain background.
小鼠胚胎干细胞（mESCs）是基因工程、基于干细胞的治疗方法开发以及多能性和早期谱系定向基础研究的关键工具。然而，直到最近，能产生有生殖系能力的胚胎干细胞系的成功衍生仍局限于少数近交小鼠品系。最近，胚胎干细胞生物学领域取得了显著进展，特别是在mESCs来源的上胚层多能性维持方面。在此，我们描述了一种从任何小鼠品系高效衍生有生殖系能力的mESCs的方案，包括以前被认为不允许的品系。我们提供了一种通常适用于大多数近交品系的方案，以及一种针对不允许品系的变体方案。通过使用该方案，mESCs可在3周内衍生出来，并在另外12周后进行全面表征，效率高达90%，且适用于任何品系背景。
