The transfer of human artificial chromosomes via cryopreserved microcells
The transfer of human artificial chromosomes via cryopreserved microcells
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DOI：
10.1007/s10616-013-9548-4
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发表时间：
2013-10-01
期刊：
影响因子：
2.2
通讯作者：
Oshimura, Mitsuo
中科院分区：
文献类型：
--
作者：
Uno, Narumi;Uno, Katsuhiro;Oshimura, Mitsuo
微细胞介导的染色体转移（MMCT）技术能够将单个完整的哺乳动物染色体或兆碱基大小的染色体片段从供体细胞转移到受体细胞。常规的MMCT方法在微细胞纯化后立即进行。微细胞的分离和受体细胞的制备的时机是非常重要的。因此，现成的微单元可以改进和简化MMCT的过程。在此，我们建立了一种在-80 A ℃下储存微细胞的冷冻保存方法，并就MMCT的效率和转移到人HT 1080细胞的人人工染色体（HAC）的稳定性将这些细胞与常规（立即）制备的细胞进行了比较。通过FISH分析证实了微细胞杂交中的HAC转移。两种方法的染色体转移效率和HAC的留存率无显著性差异。因此，即用型微细胞的冷冻保存为MMCT提供了改进和简化的方案。
Microcell-mediated chromosome transfer (MMCT) technology enables a single and intact mammalian chromosome or megabase-sized chromosome fragments to be transferred from donor to recipient cells. The conventional MMCT method is performed immediately after the purification of microcells. The timing of the isolation of microcells and the preparation of recipient cells is very important. Thus, ready-made microcells can improve and simplify the process of MMCT. Here, we established a cryopreservation method to store microcells at -80 A degrees C, and compared these cells with conventionally- (immediately-) prepared cells with respect to the efficiency of MMCT and the stability of a human artificial chromosome (HAC) transferred to human HT1080 cells. The HAC transfer in microcell hybrids was confirmed by FISH analysis. There was no significant difference between the two methods regarding chromosome transfer efficiency and the retention rate of HAC. Thus, cryopreservation of ready-to-use microcells provides an improved and simplified protocol for MMCT.