Study on LipofectinRM mediated murine SCF genetr ansduction into umbilical blood cells / 中国生化药物杂志

ABSTRACT

Purpose　The aim is to study the transduction of m urine stem cell factor(SCF) into umbilical blood cells by LipofectinRMmediated and expression in them.Methods　Murine SCF cDNA encoding extra cellu lar domain was isolated using PCR from plasmid pRC/CMV(containing SCF gene), and then recombined into the expression vector pcDNA3,and transferred into enrichme nt cultural umbilical blood cells. Semi-quantitative RT-PCR was used to examin e the expression on mRNA level.And the effects of supernatant of transfected umb ilical blood cells were investigated alone or in coordinate with GM-CSF by colo ny formation test of human bone marrow cells in semisolid culture.Results 　SCF mRNA was expressed in transfected umbilical blood cells.The su pernat ant of transfected umbilical blood cells could increase the number of CFU-GM, s ynergizing with GM-CSF.Conclusion　The supernatant of umbil ical b lood cells transfected with vector containing mSCF gene can stimulate the colony formation of human bone marrow cells in combination with GM-CSF.