A combined epigenetic and non-genetic approach for reprogramming human somatic cells.
A combined epigenetic and non-genetic approach for reprogramming human somatic cells.
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DOI：
10.1371/journal.pone.0012297
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发表时间：
2010-08-19
期刊：
影响因子：
3.7
通讯作者：
Sidhu KS
中科院分区：
文献类型：
--
作者：
Han J;Sachdev PS;Sidhu KS
不同程度的体细胞重编程已被报道使用不同的方法。然而，这通常伴随着外源材料的使用，整体重编程效率一直很低。化学物质和小分子已被用于改善体细胞核移植（SCNT）和诱导多能干细胞（iPS）生成过程中的重编程过程。我们在此报告了首次将表观遗传学和非遗传学相结合的方法应用于体细胞重编程，即DNA甲基转移酶（DNMT）和组蛋白去乙酰化酶（HDAC）抑制剂和人类胚胎干细胞（hESC）提取物。当体细胞在暴露于hESC （MEL1）提取物之前用这些抑制剂预处理时，形态学分析显示hESC样集落形成率高于未预处理的体细胞。定量PCR （qPCR）结果表明，与体细胞或单独处理hESC提取物相比，多能性基因表达上调。预处理体细胞甲基化和乙酰化水平的总体变化表明，细胞的表观遗传状态对hESC提取物诱导的重编程效率有影响。KnockOutserum replacement （KOSR™）medium （KO-SR）在诱导多能性基因表达方面发挥了积极作用。hESC提取物可能是一种无需引入外源材料就能对体细胞进行重编程的替代方法。体细胞的表观遗传预处理可以提高重编程过程的效率。在分化条件下，重编程细胞表现出向神经元分化的能力，这表明虽然重编程不能完全实现，但重编程后的细胞可以进行转分化。
Reprogramming of somatic cells to different extents has been reported using different methods. However, this is normally accompanied by the use of exogenous materials, and the overall reprogramming efficiency has been low. Chemicals and small molecules have been used to improve the reprogramming process during somatic cell nuclear transfer (SCNT) and induced pluripotent stem (iPS) cell generation. We report here the first application of a combined epigenetic and non-genetic approach for reprogramming somatic cells, i.e., DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors, and human embryonic stem cell (hESC) extracts. When somatic cells were pretreated with these inhibitors before exposure to hESC (MEL1) extracts, morphological analysis revealed a higher rate of hESC-like colony formation than without pretreatment. Quantitative PCR (qPCR) demonstrated that pluripotency genes were upregulated when compared to those of somatic cells or treated with hESC extracts alone. Overall changes in methylation and acetylation levels of pretreated somatic cells suggests that epigenetic states of the cells have an effect on reprogramming efficiency induced by hESC extracts. KnockOutserum replacement (KOSR™) medium (KO-SR) played a positive role in inducing expression of the pluripotency genes. hESC extracts could be an alternative approach to reprogram somatic cells without introducing exogenous materials. The epigenetic pre-treatment of somatic cells could be used to improve the efficiency of reprogramming process. Under differentiation conditions, the reprogrammed cells exhibited differentiation ability into neurons suggesting that, although fully reprogramming was not achieved, the cells could be transdifferentiated after reprogramming.
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