[The enhancing effects of IL-3 gene transfected murine bone marrow stromal cells on hematopoiesis under control of doxycycline].
To study enhancing effect of IL-3 gene transfected bone marrow stromal cell which can be induced by doxycycline (Dox) to express IL-3 cytokine on the proliferation and differentiation of hematopoietic stem cell, retrovirus vector system contained mIL-3 cDNA was established and bone marrow stromal cell line was transfected, and obtained QXMSC1Tet-on-IL-3, in which expression level of IL-3 can be modulated by Dox. The activities of IL-3 were measured under different Dox concentrations. The numbers of hematopoietic progenitors (CFU-GM, CFU-E, CFU-GEMM and LTC-IC) were measured and the capacity of QXMSC1Tet-on-IL-3 sustaining hematopoietic progenitor cell growth was evaluated. The results showed that IL-3 gene transfected stromal cell line QXMSC1-Tet-on + IL-3 expressed high concentration of IL-3 in vitro under control of Dox. The supernatant of QXMSC1-Tet-on + IL-3 was able to increase the number of CFU-GM, CFU-E and CFU-GEMM. The total numbers of nucleated cells and long-term cultured colonies increased in LTC-IC assay. It is concluded that in the culture of QXMSC1-Tet-on + IL-3 cells, Dox actually enhanced IL-3 expression, and thus augmented the proliferation and differentiation of hematopoietic stem/progenitor cells in vitro.
Jiang
Ji-Yang
JY
Department of Immunology, Peking University Health Science Center, Beijing 100083, China.
Li
Ai-Ling
AL
Xie
Shu-Sheng
SS
D000818:Animals; D001854:Bone Marrow Cells / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D004318:Doxycycline / Q000494:pharmacology*; D006410:Hematopoiesis / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D007377:Interleukin-3 / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D017154:Stromal Cells; D014162:Transfection
D007377:Interleukin-3; D004318:Doxycycline