Kluge N K, Ostertag W, Fusco A, Pennie S, Greenberger J S
Leuk Res. 1986;10(2):187-93. doi: 10.1016/0145-2126(86)90041-x.
The only factor-dependent or factor-independent hemopoietic murine stem cell lines which can be permanently maintained in vitro are cell lines which originate from bone marrow cultures of congenic mice differing in the Fv-2 locus infected with Friend spleen focus forming virus (F-SFFV) in conjunction with either Rauscher or Friend helper virus (R-MuLV or F-MuLV). We determined the viral state of these cells by restriction enzyme analysis, measurement of SFFV and MuLV related RNA, immunoprecipitation analysis of viral related proteins and biological activity to test whether integration and expression of R-MuLV or F-SFFV are obligatory. All stem cell lines (SUT, JUT, 416B) showed expression of MuLV coded proteins or virus but SFFV was not found in these cell lines. A two-fold difference of RNA hybridizing with SFFV-specific cDNAs observed in stem cells of Fv-2rr and of Fv-2ss genotype is in agreement with data published earlier.
唯一能够在体外永久维持的依赖因子或不依赖因子的造血小鼠干细胞系,是那些源自Fv-2位点不同的同基因小鼠骨髓培养物的细胞系,这些培养物感染了弗氏脾脏病灶形成病毒(F-SFFV),并与劳舍尔或弗氏辅助病毒(R-MuLV或F-MuLV)联合使用。我们通过限制性内切酶分析、SFFV和MuLV相关RNA的测量、病毒相关蛋白的免疫沉淀分析以及生物活性来确定这些细胞的病毒状态,以测试R-MuLV或F-SFFV的整合和表达是否是必需的。所有干细胞系(SUT、JUT、416B)均显示出MuLV编码蛋白或病毒的表达,但在这些细胞系中未发现SFFV。在Fv-2rr和Fv-2ss基因型的干细胞中观察到与SFFV特异性cDNA杂交的RNA有两倍差异,这与早期发表的数据一致。