...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular reproduction and development >Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris Altaica).
【24h】

Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris Altaica).

机译：西伯利亚虎（Panthera tigris Altaica）体外培养的皮肤成纤维细胞的细胞周期分析和种间核转移。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The present study was conducted to examine the effect of cell culture conditions, antioxidants, protease inhibitors (PI), and different levels of dimethylsulfoxide (DMSO) for the promotion of synchronization of different cell cycles of Siberian tiger skin fibroblasts. We also compared the ability of somatic cell nuclei of the Siberian tiger in pig cytoplasts and to support early development after reconstruction. Cell cycle synchronization between nuclear donor and recipient cells is considered to be one of the most crucial factors for successful cloning. Five experiments were performed each with a one-way completely randomized design involving three replicates of all treatments. Least significant difference (LSD) was used to determine variation among treatment groups. Experiment I focused in the effects of cycling, serum starved and fully confluent stages of Siberian tiger cells on different cell cycles. In Experiment II, the effects of different antioxidants like beta-Mercaptoethanol (beta-ME, 10 microM), cysteine (2 mM), and glutathione (2 mM) were examined after cells were fully confluent without serum starvation for 4 hr. In Experiment III, three PI, namely 6-dimethylaminopurine (6-DMAP, 2 mM), cycloheximide (7.5 microg/ml) and cytochalasin B (7.5 microg/ml) were used in the sane manner as in Experiment II. In Experiment IV, different levels of DMSO at 0%, 0.5%, 1.0%, and 2.5% were tested on different cell cycle stages of Siberian tiger examined by Flowcytometry (FACS). In Experiment I, 67.2% of the Siberian tiger skin fibroblasts reached the G0/G1 stage (2C DNA content) in fully confluent conditions which was more than the cycling (49.8%) and serum starved (SS) medium (65.5%; P < 0.05). Among the chemically treated group, glutathione (72.6%) and cycloheximide (71.3%) had little bit better results for the synchronization of G0 + G1 phases than serum starved and fully confluent. After nuclear transfer we did not see any significant differences on the development of tiger-porcine reconstructed embryos at cycling, SS and fully confluent. Data indicate that prolonged culture of cells in the absence of serum as well as using different chemicals for this experiment does not imply a shift in the percentage of cells that enter G0/G1 and that confluency is sufficient to induce quiescence. This finding can be beneficial in nuclear transfer programs in Siberian tiger, because there are negative effects, such as apoptosis associated with serum starvation.
机译：进行本研究以检查细胞培养条件，抗氧化剂，蛋白酶抑制剂（PI）和不同水平的二甲基亚砜（DMSO）对促进西伯利亚虎皮成纤维细胞不同细胞周期同步的影响。我们还比较了猪细胞质中西伯利亚虎体细胞核的能力以及重建后支持早期发育的能力。核供体和受体细胞之间的细胞周期同步被认为是成功克隆的最关键因素之一。用单向完全随机设计进行了五个实验，涉及所有处理的三个重复。最小显着差异（LSD）用于确定治疗组之间的差异。实验一集中在西伯利亚虎细胞的循环，血清饥饿和完全融合阶段对不同细胞周期的影响。在实验II中，在细胞完全汇合且无血清饥饿4小时后，检查了不同的抗氧化剂如β-巯基乙醇（β-ME，10 microM），半胱氨酸（2 mM）和谷胱甘肽（2 mM）的作用。在实验III中，以与实验II中相同的方式使用三种PI，即6-二甲基氨基嘌呤（6-DMAP，2mM），环己酰亚胺（7.5μg/ ml）和细胞松弛素B（7.5μg/ ml）。在实验IV中，通过流式细胞术（FACS）在西伯利亚虎的不同细胞周期阶段测试了0％，0.5％，1.0％和2.5％的DMSO含量不同。在实验一中，西伯利亚虎皮成纤维细胞中有67.2％在完全融合的条件下达到了G0 / G1阶段（2C DNA含量），高于循环（49.8％）和血清饥饿（SS）培养基（65.5％; P < 0.05）。在化学治疗组中，谷胱甘肽（72.6％）和环己酰亚胺（71.3％）的G0 + G1相同步结果好于血清饥饿和完全融合。核移植后，在循环，SS和完全融合时，在虎猪重建胚胎的发育上没有发现任何显着差异。数据表明，在无血清的情况下延长细胞的培养时间以及在本实验中使用不同的化学药品并不意味着进入G0 / G1的细胞百分比发生了变化，并且汇合足以诱导静止。这一发现在西伯利亚虎的核移植计划中可能是有益的，因为它具有负面影响，例如与血清饥饿相关的细胞凋亡。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号