Derivation of stable embryonic stem cell-like, but transcriptionally heterogenous, induced pluripotent stem cells from non-permissive mouse strains.
Derivation of stable embryonic stem cell-like, but transcriptionally heterogenous, induced pluripotent stem cells from non-permissive mouse strains.
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DOI：
10.1007/s00335-020-09849-x
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发表时间：
2020-12
期刊：
影响因子：
2.5
通讯作者：
Threadgill, David W.
中科院分区：
文献类型：
--
作者：
Garbutt, Tiffany A.;Konganti, Kranti;Konneker, Thomas;Hillhouse, Andrew;Phelps, Drake;Jones, Alexis;Aylor, David;Threadgill, David W.
众所周知，遗传背景在获得多能胚胎干细胞(ESC)的能力中发挥着作用，这种特征被称为迁移性。此前，我们已经证明，在重新编程三天后，通过添加添加GSK3B和MEK抑制剂的基于血清的培养液，可以很容易地从不允许的小鼠品系中获得诱导多能干细胞(IPSC)。在这里，我们描述了从不允许的小鼠品系中衍生出第二种类型的IPSC菌落，这种菌落可以稳定地维持在不依赖于2IS介质的情况下。由此产生的细胞表现出转录异质性，类似于在ESC中观察到的，来自于含有补充白血病抑制因子的常规血清的允许遗传背景。然而，与以前报道排他性亚群的研究不同，我们观察到单纯和启动的细胞表面标记的排他性和同时表达。在这里，我们探索了在2IS存在的情况下多能性的变化，并表征了异源亚群，以确定它们的多能性状态和在来自非允许的NOD/ShiLtd J株的异源IPSCs中的作用。我们的结论是，异质性是干细胞的一种自然产生的、必要的品质，允许维持多能性。这项研究进一步证明了2IS重编程技术的有效性。这也是第一次从不允许的NOD/ShiLtd J和WSB/EIJ菌株中获得稳定的ESC样干细胞的研究，使这些和类似的不允许的小鼠品系和物种的多能性研究变得更容易和更广泛。
Genetic background is known to play a role in the ability to derive pluripotent, embryonic stem cells (ESC), a trait referred to as permissiveness. Previously we have demonstrated that induced pluripotent stem cells (iPSC) can be readily derived from non-permissive mouse strains by addition of serum-based media supplemented with GSK3B and MEK inhibitors, termed 2iS media, three days into reprogramming. Here we describe the derivation of second type of iPSC colony from non-permissive mouse strains that can be stably maintained independently of 2iS media. The resulting cells display transcriptional heterogeneity similar to that observed in ESC from permissive genetic backgrounds derived in conventional serum containing media supplemented with leukemia inhibitor factor. However, unlike previous studies that report exclusive subpopulations, we observe both exclusive and simultaneous expression of naive and primed cell surface markers. Herein, we explore shifts in pluripotency in the presence of 2iS and characterize heterogenous subpopulations to determine their pluripotent state and role in heterogenous iPSCs derived from the non-permissive NOD/ShiLtJ strain. We conclude that heterogeneity is a naturally occurring, necessary quality of stem cells that allows for the maintenance of pluripotency. This study further demonstrates the efficacy of the 2iS reprogramming technique. It is also the first study to derive stable ESC-like stem cells from the non-permissive NOD/ShiLtJ and WSB/EiJ strains, enabling easier and broader research possibilities into pluripotency for these and similar non-permissive mouse strains and species.
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影响因子：
21.3
作者：
Boroviak T;Loos R;Bertone P;Smith A;Nichols J
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