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目的：拓展造血干细胞因子（ＳＣＦ）的研究和基因治疗途径。方法：利用基因重组技术，构建了编码可溶性人ＳＣＦ基因的逆转录病毒载体ｐＬＸＳＮ－ＳＣＦ，应用脂质体ｌｉｐｏｆｅｃｔｉｎ基因转移法将重组质粒导入Ψ２和ＰＡ３１７病毒包装细胞，经Ｇ４１８选择性培养基筛选，获得产重组病毒的包装细胞ＰＡ３１７／ＳＣＦ，其病毒效价为（２．４～８．５）×１０５ＣＦＵ／ｍｌ，继而感染人造血干／祖细胞。应用多聚酶链反应、ＡＰＡＡＰ免疫组化染色和化学发光－直接酶标法检测人ＳＣＦ基因在细胞中转移和表达。结果：逆转录病毒载体介导的ｒｈＳＣＦ基因在人造血干／祖细胞中获得有效转移和表达。结论：为进一步研究ＳＣＦ的生物学特性及开展干细胞移植等提供了一定的途径。
Objective: To explore the research and gene therapy approaches of hematopoietic stem cell factor (SCF). Methods: The retroviral vector pLXSN-SCF encoding the soluble human SCF gene was constructed by gene recombination technique. The recombinant plasmids were introduced into the packaging cells of Ψ2 and PA317 virus by lipofectin gene transfer method and screened by G418 selective medium. PA317 / SCF, a packaging cell producing recombinant virus, was obtained and its virus titer was (2.4 ~ 8.5) × 105CFU / ml, which in turn infected human hematopoietic stem / progenitor cells. Polymerase chain reaction, APAAP immunohistochemical staining and chemiluminescence-direct enzyme-linked immunosorbent assay were used to detect the metastasis and expression of human SCF gene. Results: The retroviral vector-mediated rhSCF gene was efficiently transferred and expressed in human hematopoietic stem / progenitor cells. Conclusion: It provides a certain way to further study the biological characteristics of SCF and carry out stem cell transplantation.