Reprogramming events of mammalian somatic cells induced by Xenopus laevis egg extracts
Reprogramming events of mammalian somatic cells induced by Xenopus laevis egg extracts
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DOI：
10.1002/mrd.20691
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发表时间：
2007-10-01
影响因子：
2.5
通讯作者：
Imai, Hiroshi
中科院分区：
文献类型：
--
作者：
Miyamoto, Kei;Furusawa, Tadashi;Imai, Hiroshi
已知核移植后，分化的细胞可以在卵母细胞胞质中重编程为未分化状态。近年来，有研究表明非洲爪蟾卵提取物具有对哺乳动物体细胞进行重编程的能力。尚未阐明爪蟾卵提取物处理后和提取物处理的细胞的细胞培养后哺乳动物细胞的重编程事件。在这个实验中，我们研究了爪蟾卵提取物处理后可逆透化或非透化猪成纤维细胞中的重编程事件。非洲爪蟾卵特异性组蛋白134组装在猪染色质上，核核纤层蛋白LIII被掺入细胞核中。组蛋白H3在提取物处理的细胞中的赖氨酸9的脱乙酰化在nonpermeabilized细胞中被检测到，这表明即使在nonpermeabilized细胞中也可以诱导重编程的一部分。培养提取物处理的细胞后，细胞开始表达多能标记基因如POU 5 F1（OCT 4）和SOX 2，并形成集落。在用OCT 4-EGFP构建体转化的牛成纤维细胞中也证实了提取物处理的细胞中OCT 4基因的再活化。这些结果表明，哺乳动物细胞的细胞核可以部分重编程为胚胎状态的爪蟾卵提取物和重塑细胞部分去分化后的细胞培养。利用卵提取物的系统可能有助于了解哺乳动物体细胞核移植后去分化和重编程的机制和过程。
It is known that differentiated cells can be reprogrammed to an undifferentiated state in oocyte cytoplasm after nuclear transfer. Recently, some reports suggested that Xenopus egg extracts have the ability to reprogram mammalian somatic cells. Reprogramming events of mammalian cells after Xenopus egg extract treatment and after cell culture of extract-treated cells have not been elucidated. In this experiment, we examined reprogramming events in reversibly permeabilized or nonpermeabilized porcine fibroblast cells after Xenopus egg extract treatment. The Xenopus egg-specific histone 134 was assembled on porcine chromatin and nuclear lamin LIII was incorporated into nuclei. Deacetylation of histone H3 at lysine 9 in extract-treated cells was detected in nonpermeabilized cells, suggesting that a part of reprogramming may be induced even in nonpermeabilized cells. Following culture of extract-treated cells, the cells began to express the pluripotent marker genes such as POU5F1 (OCT4) and SOX2 and to form colonies. Reactivation of the OCT4 gene in extract-treated cells was also confirmed in bovine fibroblasts transformed with an OCT4-EGFP construct. These results suggest that nuclei of mammalian cells can be partially reprogrammed to an embryonic state by Xenopus egg extracts and the remodeled cells partly dedifferentiate after cell culture. A system using egg extracts may be useful for understanding the mechanisms and processes of dedifferentiation and reprogramming of mammalian somatic cells after nuclear transfer.