| 家蚕BmN细胞S期同步化体系建立 |
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| 引用本文： | 黄蕾,江千令,阚云超,李丹丹.家蚕BmN细胞S期同步化体系建立[J].中国农学通报,2014,30(2):39-44. |
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| 作者姓名： | 黄蕾 江千令 阚云超 李丹丹 |
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| 作者单位： | 1. 2. 南阳师范学院 |
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| 基金项目： | 家蚕非编码RNA Bm-15在Notch信号通路及发育过程中的功能研究;调控家蚕发育非编码RNA（non-coding RNA，ncRNA）的功能解析 |
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| 摘 要： | 细胞S期为DNA复制期，探索细胞S期的同步化将为研究各种调控因子对DNA复制、细胞分裂和增殖的影响奠定基础。本研究以家蚕BmN细胞为研究对象，采用脱氧胸苷(TdR)双阻断、脱氧胸苷-诺考达唑(TdR-Nocodazole)双阻断及羟基脲(HU)法，探索BmN细胞S期同步化的最佳体系。结果发现，TdR双阻断法处理对BmN细胞S期的同步化效率为67.2%，TdR-Nocodazole双阻断法处理为69.7%，HU处理对S期同步化效果最好，达90%以上。进一步对HU处理后不同释放时间点的细胞进行研究发现，1 mmol/mL HU处理后释放7 h时BmN细胞S期同步化效率最高，达94.9%。通过不同处理方法研究家蚕BmN细胞的S期同步化效果，发现1 mmol/mL的HU处理16 h，然后于正常培养基中释放7 h，可有效的将BmN细胞同步化在S期，且该处理方法简单，易于操作。
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| 关 键 词： | 品种 品种 |
| 收稿时间： | 2013/6/13 0:00:00 |
| 修稿时间： | 2013/7/11 0:00:00 |
Establishment of S Phage Synchronization system for BmN Cells |
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| Huang Lei;Jiang Qianling;Kan Yunchao;Li Dandan.Establishment of S Phage Synchronization system for BmN Cells[J].Chinese Agricultural Science Bulletin,2014,30(2):39-44. |
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| Authors: | Huang Lei;Jiang Qianling;Kan Yunchao;Li Dandan |
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| Institution: | Huang Lei;Jiang Qianling;Kan Yunchao;Li Dandan;China-UK-NYNU-RRes Joint Libratory of insect biology/ Nanyang Normal University; |
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| Abstract: | OBJECTIVE] S phage is important in cell cycle, during which DNA replication occurs, some anti-cancer drugs are designed to block the replication of DNA and plays efficacy. BmN cells are widely used in the research of silkworm, studies on the synchronization of S phase of BmN cells will build foundation on the researches of regulation of cell proliferation, division and apoptosis. METHOD] Three methods (TdR double occlusion, TdR-Nocodazole double occlusion and hydroxyurea occlusion) were used to study the synchronization effect on S phage of BmN cells. RESULTS] Results showed that TdR double occlusion had S phage sychronization efficiency of 67.2%, TdR-Nocodazole double occlusion was up to 69.7%, while hydroxyurea had the best sychronization efficiency with percentage more than 90%, which was similar to the results found in Sf9 cells and drosophila cells. Then we further analyzed the detailed conditions of HU occlusion, we found that after 16h treatment by 1mmol/mL HU then being released in normal medium with 7h, the S phase cells of BmN were up to 94.9%. CONCLUSION] Different methods were used to study the S phage synchronization efficiecy of BmN cells, results showed that treatment of 1mmol/mL HU with 16h and then being released in normal medium with 7h was the best synchronization method for BmN cells with simple and easy operation. |
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| Keywords: | hydroxyurea |
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