| 制作方法Making method |
We transplanted human CD34+ hematopoietic stem cells into the liver of new-born recipient NOD/SCID/Il2rgnull mice (NOG mice). Afer 10–12 weeks post-transplantation, the humanized mice (n=6) were intraperitoneally infected with the CCR5-tropic HIV-1JR-CSF. We divided the six mice into two groups; respectively 2 weeks post infection (wpi) and 15 wpiNOD/SCID/Il2rgnull mice (NOG mice) were obtained from the Central Institute for Experimental Animals (Kawasaki, Japan)39. Human CD34+ hematopoietic stem cells were isolated from human fetal liver kindly provided by Dr. Dong Sung An (University of Los Angeles, USA). We generated humanized mice (NOG-hCD34 mice) as described previously40, 41. HIV-1JR-CSF viral solution was prepared as previously described42. Briefy, pJR-CSF, an infectious molecular clone of HIV-1JR-CSF43, 44, was transfected into 293T cells. At 48 h post-transfection, the culture supernatant was harvested, centrifuged, and then fltered through a 0.45-μm-pore-size flter to produce virus solutions. Viral solution (1,500 infectious units [IU]) was intraperitoneally injected into the NOG-hCD34 mice. |
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