Gruber H E, Finley K D, Hershberg R M, Katzman S S, Laikind P K, Seegmiller J E, Friedmann T, Yee J K, Jolly D J
Science. 1985 Nov 29;230(4729):1057-61. doi: 10.1126/science.3864246.
The transfer of the human gene for hypoxanthine phosphoribosyltransferase (HPRT) into human bone marrow cells was accomplished by use of a retroviral vector. The cells were infected in vitro with a replication-incompetent murine retroviral vector that carried and expressed a mutant HPRT complementary DNA. The infected cells were superinfected with a helper virus and maintained in long-term culture. The production of progeny HPRT virus by the bone marrow cells was demonstrated with a colony formation assay on cultured HPRT-deficient, ouabain-resistant murine fibroblasts. Hematopoietic progenitor cells able to form colonies of granulocytes or macrophages (or both) in semisolid medium in the presence of colony stimulating factor were present in the nonadherent cell population. Colony forming units cloned in agar and subsequently cultured in liquid medium produced progeny HPRT virus, indicating infection of this class of hematopoietic progenitor cell.
通过使用逆转录病毒载体，将人类次黄嘌呤磷酸核糖基转移酶（HPRT）基因转移到人类骨髓细胞中。这些细胞在体外被一种无复制能力的鼠逆转录病毒载体感染，该载体携带并表达突变的HPRT互补DNA。感染的细胞被辅助病毒超感染并维持长期培养。通过对培养的HPRT缺陷、哇巴因抗性鼠成纤维细胞进行集落形成试验，证明骨髓细胞产生了子代HPRT病毒。在非贴壁细胞群体中存在造血祖细胞，它们在集落刺激因子存在的情况下，能够在半固体培养基中形成粒细胞或巨噬细胞（或两者）的集落。在琼脂中克隆并随后在液体培养基中培养的集落形成单位产生了子代HPRT病毒，表明这类造血祖细胞受到了感染。
