Hesdorffer C, Ward M, Markowitz D, Bank A
Columbia University College of Physicians and Surgeons, Department of Medicine, New York, NY 10032.
DNA Cell Biol. 1990 Dec;9(10):717-23. doi: 10.1089/dna.1990.9.717.
Irradiated mice were transplanted with cells containing a foreign gene to evaluate gene transfer and expression as a model for gene therapy. Using a uniquely safe and efficient retroviral packaging line developed in this laboratory (GP + E86), we demonstrate efficient and safe long-term transfer of the neomycin resistance (neoR) gene into mice. By infusing cells obtained from spleen colonies of primary post-transplant mice marked with the neoR gene into irradiated recipients, secondary and tertiary generations of recipient mice were produced. There was very low reconstitution activity of single stem cells in these successive generations of mice. We conclude that many more than one stem cell is necessary for successful long-term bone marrow transplantation in mice, presumably as a result of the relatively low frequency of stem cell cycling.
对经辐照的小鼠进行含有外源基因的细胞移植，以评估基因转移和表达情况，作为基因治疗的一种模型。利用本实验室开发的一种独特安全且高效的逆转录病毒包装细胞系（GP + E86），我们证明了新霉素抗性（neoR）基因能有效且安全地长期转移到小鼠体内。通过将来自用neoR基因标记的移植后初代小鼠脾脏集落的细胞注入经辐照的受体小鼠体内，产生了受体小鼠的第二代和第三代。在这些连续几代的小鼠中，单个干细胞的重建活性非常低。我们得出结论，在小鼠中成功进行长期骨髓移植需要不止一个干细胞，这可能是由于干细胞循环频率相对较低的结果。
