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目的 探索将外源性ＩＬ－３ｃＤＮＡ导入骨髓基质细胞的可能性。方法 将小鼠ＩＬ－３ｃＤＮＡ克隆到逆转录病毒载体（ｐＬＸ－ＳＮ）上，然后将重组的ＩＬ－３ ｃＤＮＡ用脂质体转染到包装细胞ＰＡ３１７上，Ｇ４１８筛选后得到抗性克隆，将含病毒的上清液转染骨髓基质细胞。用自行设计的引物，检测ｐＬＸＳＮ上的ＮｅｏＲ基因及转入的ＩＬ－３基因。同时观察了转染ｐＬＸＳＮ／ＩＬ－３的基质细胞培养上清液对小鼠骨髓 ＣＦＵ－ＧＭ形成的影响，并测定其对造血因子依赖株 ＮＦＳ－６０细胞增殖活性的影响。结果 将含重组子 ｐＬＸＳＮ－ＩＬ－３和空载体ｐＬＸＳＮ的病毒培养上清感染 ＢＭＳ，前者用 ＰＣＲ扩增出５００ ｂｐ和１７０ ｂｐ（ＮｅｏＲ）的条带，后者仅扩增出 １７０ ｂｐ。转染ｐＬＸＳＮ／ＩＬ－３的基质细胞上清液有促进小鼠骨髓ＣＦＵ－ＧＭ增殖的作用，与空载体组及未转染组比较，有非常显著性差异（Ｐ＜０．０１）。结论 外源性的ＩＬ－３基因及ｐＬＸＳＮ上的ＮｅｏＲ已整合到骨髓基质细胞基因组ＤＮＡ中，并促进造血细胞增殖。
Objective To explore the possibility of introducing exogenous IL-3 cDNA into bone marrow stromal cells. Methods Mouse IL-3 cDNA was cloned into retroviral vector (pLX-SN). The recombinant IL-3 cDNA was transfected into packaging cell PA317 by lipofectamine 2000. After selection with G418, The virus supernatant was transfected into bone marrow stromal cells. Using self-designed primers, the NeoR gene on pLXSN and the transfected IL-3 gene were detected. At the same time, we observed the effect of culture supernatants transfected with pLXSN / IL-3 on the CFU-GM formation in mouse bone marrow and the effect on the proliferation of the hematopoietic line-dependent NFS-60 cells. Results The virus supernatant containing pLXSN-IL-3 and empty vector pLXSN was infected with BMS. The former amplified bands of 500 bp and 170 bp (NeoR) by PCR, and the latter amplified only 170 bp. The supernatant of stromal cells transfected with pLXSN / IL-3 could promote the proliferation of mouse bone marrow CFU-GM. Compared with the empty vector group and the untransfected group, there was a significant difference (P <0.01). Conclusion The exogenous IL-3 gene and NeoR on pLXSN have been integrated into genomic DNA of bone marrow stromal cells and promote hematopoietic cell proliferation.