爪蟾卵母细胞抽提物可诱导牛胎儿成纤维细胞发生部分重编程

doi:10.11843/j.issn.0366-6964.2015.09.009

摘要/Abstract

摘要：

应用非洲爪蟾卵母细胞抽提物处理和牛胎儿成纤维细胞（Japanese Black cattle fetal fibroblasts，JBCFF），观察处理前后细胞的形态学变化，同时用免疫荧光染色法检测组蛋白的乙酰化状态和OCT4蛋白的表达，并对其多能性标志基因的mRNA表达水平进行定量分析。结果表明，与未处理细胞相比，经抽提物处理和牛胎儿成纤维细胞组蛋白H3K9乙酰化程度与未处理组无显著差异；培养5~6 d后细胞聚集形成“克隆簇”中碱性磷酸酶和Oct4蛋白染色阳性；同时也检测到Oct4和Nanog基因在其中的表达，而Sox2基因未见表达；且Oct4、Nanog基因表达量随处理后细胞培养时间的延长（4、5、6 d）而呈依次上升趋势。可见，爪蟾卵母细胞抽提物能诱导和牛胎儿成纤维细胞发生部分重编程，恢复其发育全能性，这对牛诱导性干细胞制备方法的探索和体细胞克隆及转基因克隆牛效率的提高具有一定的借鉴意义。

Abstract:

In order to reprogram the Japanese Black cattle fetal fibroblasts(JBCFF)，the Xenopus laevis oocyte extracts was used to incubation with the cells in present study.After 24 hours of culture in medium，there was no significant difference in acetylation of histone H3K9 between the JBCFF treated with extracts and the cells untreated by immunofluorescence assay.The well-defined colony structures of JBCFF treated with extracts were formed with the continuous culture for 5-6 d and the alkaline phosphatase staining of the cells colony was positive.Similarly，the OCT4 protein was detected in the cells colony.Additionally，Oct4 and Nanog mRNA expression，two pluripotency marker genes，were detected in those cells by RT-PCR，but Sox2 gene expression was not detected.Furthermore，qPCR results showed that the expression levels of Oct4 and Nanog genes in colony cells increased with days of culture(4，5，6 d ) after extracts treatment.In conclusion，the Xenopus laevis oocyte extracts could induce the partial reprogramming of the JBCFF into a low differentiated state.

中图分类号:

S823.2

杜卫华，范宗兴，王皓宇，郝海生，刘岩，赵学明，秦彤，朱化彬. 爪蟾卵母细胞抽提物可诱导牛胎儿成纤维细胞发生部分重编程[J]. 畜牧兽医学报, 2015, 46(9): 1549-1556.

DU Wei-hua，FAN Zong-xing，WANG Hao-yu，HAO Hai-sheng，LIU Yan，ZHAO Xue-ming，QIN Tong，ZHU Hua-bin. Reprogramming of Japanese Black Cattle Fetal Fibroblasts Treated with Xenopus laevis Oocytes Extracts[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(9): 1549-1556.

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https://www.xmsyxb.com/CN/Y2015/V46/I9/1549

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