Derivation of LIF-Independent Embryonic Stem Cells Using Inducible OCT4 Expression.
Derivation of LIF-Independent Embryonic Stem Cells Using Inducible OCT4 Expression.
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DOI：
10.1007/978-1-0716-0301-7_13
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发表时间：
2020
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通讯作者：
Benjamin L. Kidder
中科院分区：
文献类型：
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作者：
Benjamin L. Kidder
多能性小鼠胚胎干细胞（ES细胞）来源于植入前胚胎的内细胞团（ICM），在存在外部信号白血病抑制因子（LIF）的情况下能够无限期地自我更新，通过抑制GSK 3激活Wnt信号传导，并抑制MAP激酶/ERK激酶信号传导。OCT 4转录因子在多能细胞中高度表达，是多能状态的中心转录调节因子。在这里，我们描述了一种使用诱导型OCT 4（iOCT 4）ES细胞模型系统在LIF非依赖性和无血清培养基中培养ES细胞的方案。该方案足以在没有外部信号的情况下在限定条件下维持ES细胞体外自我更新。LIF非依赖性iOCT 4 ES细胞在诱导型OCT 4转基因失活后完全能够分化。
Pluripotent mouse embryonic stem (ES) cells, which are derived from the inner cell mass (ICM) of preimplantation stage embryos, are capable of self-renewing indefinitely in the presence of the external signal leukemia inhibitory factor (LIF), activation of Wnt signaling through inhibition of GSK3, and inhibition of MAP kinase/ERK kinase signaling. The OCT4 transcription factor is expressed highly in pluripotent cells and is a central transcriptional regulator of the pluripotent state. Here, we describe a protocol to culture ES cells in LIF-independent and serum-free media using an inducible OCT4 (iOCT4) ES cell model system. This protocol is sufficient to sustain ES cell self-renewal in vitro in defined conditions in the absence of external signals. LIF-independent iOCT4 ES cells are fully capable of differentiating following deactivation of the inducible OCT4 transgene.