Takahashi M, Furukawa T, Tanaka I, Nikkuni K, Aoki A, Kishi K, Koike T, Moriyama Y, Shibata A
First Department of Internal Medicine, Niigata University School of Medicine, Japan.
Leuk Res. 1992 Aug;16(8):761-7. doi: 10.1016/0145-2126(92)90154-y.
Our previous study demonstrated the positive relationship between the gene introduction rate into hematopoietic cell lines by electroporation and the percentage of cells in S-phase. In the present study, granulocyte-macrophage progenitor cells (CFU-C) rich marrow cell fraction were cultured in suspension with IL-3, GM-CSF and G-CSF for 4 days. The number of CFU-C were increased three times after the culture, and 3H-thymidine suicide tests of cultured cells demonstrated that the proportion of CFU-C in S-phase was increased by two to four times. The efficiency of gene transfer into CFU-C with the plasmid pMoZtk (containing the beta-galactosidase gene) by electroporation was nearly doubled by culturing marrow cells with these growth factors. These findings confirm that the introduction rate of the gene into CFU-C by electroporation is more efficient in cell populations with a higher percentage of CFU-C in S-phase.
我们之前的研究表明，通过电穿孔将基因导入造血细胞系的效率与处于S期的细胞百分比之间存在正相关关系。在本研究中，富含粒细胞-巨噬细胞祖细胞（CFU-C）的骨髓细胞部分与白细胞介素-3、粒细胞巨噬细胞集落刺激因子和粒细胞集落刺激因子在悬浮液中培养4天。培养后CFU-C的数量增加了两倍，对培养细胞进行的3H-胸腺嘧啶核苷自杀试验表明，处于S期的CFU-C比例增加了两到四倍。通过电穿孔用质粒pMoZtk（含β-半乳糖苷酶基因）将基因导入CFU-C的效率，通过用这些生长因子培养骨髓细胞几乎提高了一倍。这些发现证实，在处于S期的CFU-C百分比更高的细胞群体中，通过电穿孔将基因导入CFU-C的效率更高。
