Stability of transferred human chromosome fragments in cultured cells and in mice
Stability of transferred human chromosome fragments in cultured cells and in mice
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DOI：
10.1023/a:1026741321193
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发表时间：
2000-01-01
影响因子：
2.6
通讯作者：
Oshimura, M
中科院分区：
文献类型：
--
作者：
Shinohara, T;Tomizuka, K;Oshimura, M
利用微细胞介导的染色体转移，染色体片段是可行的基因传递载体。为了测试基因传递载体的先决条件，我们检测了人类染色体片段（hcf）在培养细胞和跨染色体小鼠（Tc）中的稳定性。将neo标记的人2号染色体（hCF(2- w23)）、11号染色体（hCF-11）和14号染色体（hCF(SC20)）片段导入TT2F小鼠ES细胞，在不选择的长期培养过程中，用FISH检测hCF的保留情况。与hCF（2-W23）和hCF-11的逐渐丢失相反，hCF（SC20）在胚胎干细胞中保持稳定超过70倍。hCF（SC20）在培养的人肿瘤细胞和鸡DT40细胞中也保持稳定。我们之前使用含有hCF（2-W23）或hCF（SC20）的胚胎干细胞产生了嵌合小鼠，随后产生了Tc小鼠。尽管hCF（2-W23）和hCF（SC20）作为附加染色体在Tc小鼠细胞中持续存在并遗传给后代，但hCF（SC20）在F1和F2小鼠中比hCF（2-W23）更稳定。本研究表明，hcf在Tc小鼠体内的稳定性随组织类型和遗传背景的不同而不同。因此，在小鼠和人类细胞中相对稳定的hCF（SC20）可能是一个有希望发展成为基因传递载体的候选者。
Chromosome fragments represent feasible gene delivery vectors with the use of microcell-mediated chromosome transfer. To test a prerequisite for a gene delivery vector, we examined the stability of human chromosome fragments (hCFs) in cultured cells and in trans-chromosomic (Tc) mice. Fragments of human chromosomes 2 (hCF(2-W23)), 11 (hCF-11) and 14 (hCF(SC20)) tagged with neo were introduced into the TT2F mouse ES cells, and retention of the hCFs was examined by FISH during long-term culture without selection. In contrast to the gradual loss of hCF(2-W23) and hCF-11, hCF(SC20) remained stable over 70 population doublings in the ES cells. The hCF(SC20) was also stable in cultured human tumor cells and chicken DT40 cells. We have previously generated chimeric mice using the ES cells harboring the hCF(2-W23) or hCF(SC20), followed by production of Tc mice. Although both the hCF(2-W23) and hCF(SC20) persisted in cells of Tc mice as an additional chromosome and were transmitted to offspring, the hCF(SC20) was more stable than the hCF(2-W23) in F1 and F2 mice. The present study shows that the stability of hCFs in Tc mice differs with tissue types and with genetic background used for successive breedings. Thus, the hCF(SC20), which was relatively stable in both mouse and human cells, may be a promising candidate for development as a gene delivery vector.