Dick J E, Magli M C, Huszar D, Phillips R A, Bernstein A
Cell. 1985 Aug;42(1):71-9. doi: 10.1016/s0092-8674(85)80102-1.
We have used the random chromosomal integration sites of retrovirus vectors as unique clonal markers to analyze cell lineage relationships within the hemopoietic stem cell hierarchy. Using a high efficiency protocol for retrovirus-mediated gene transfer, anemic W/Wv mutant mice were reconstituted with bone marrow cells infected with a NEO vector. Analysis of the DNA from bone marrow, thymus, and spleen of these reconstituted W/Wv mice indicated insertion of the vector into primitive pluripotent stem cells capable of producing both myeloid and lymphoid progeny as well as into more committed stem cells apparently restricted to either the myeloid or lymphoid lineages. The neo gene was also expressed in these mice, as they contained a variety of G418 resistant in vitro colony-forming cells. These results demonstrate high-efficiency gene transfer and expression in primitive hemopoietic stem cells and provide a direct approach for analyzing the hemopoietic stem cell hierarchy.
我们利用逆转录病毒载体的随机染色体整合位点作为独特的克隆标记，来分析造血干细胞层级结构内的细胞谱系关系。采用高效的逆转录病毒介导基因转移方案，用感染了NEO载体的骨髓细胞重建贫血的W/Wv突变小鼠。对这些重建的W/Wv小鼠的骨髓、胸腺和脾脏DNA进行分析表明，载体插入了能够产生髓系和淋巴系后代的原始多能干细胞以及显然局限于髓系或淋巴系的更定向的干细胞中。neo基因在这些小鼠中也有表达，因为它们含有多种对G418耐药的体外集落形成细胞。这些结果证明了在原始造血干细胞中的高效基因转移和表达，并为分析造血干细胞层级结构提供了直接方法。
