细胞实验
Animal study[1]
Female BALB/c mice were housed underpathogen-free conditions and maintained in a 12-hour light and 12-hour darkcycle with food and water supplied ad libitum. For Y-90 ibritumomab tiuxetantreatment, a dose of 10, 25, 50, 100 or 200 μCi of Y-90ibritumomab tiuxetan was diluted with phosphate-buffered saline (PBS), resultingin a total volume of 100μL, and was administered to mice via tail vein injection.
For mouse xenograft experiments, athymicnude mice (BALB/c Foxn1nu, 5 to 6 weeks old) were used. Inocula of 5 x 106 Karpas-422 cells, a human Bcell lymphoma cell line, in 0.1 mL of dilutedprotein gel mixture (1:1 dilution in PBS) at 4oC were injected intothe subcutaneous space on the right flanks of mice. When tumors reached 0.1 cmin diameter, 20 mice were randomized into 4 treatment groups: control, LDAonly, Y-90 ibritumomab tiuxetan only, and Y-90 ibritumomab tiuxetan with LDApretreatment. For Y-90 ibritumomab tiuxetan-treated groups, 200μCiof Y-90 ibritumomab tiuxetan was injected via the tail vein.
Tissueprocurement and immunohistochemistry
Mice were sacrificed at 6, 24, and 72 hoursand at 1 and2 weeks after treatment. Cleaned femurs and tibias were fixed in10% buffered formalin solutions and decalcified in14% EDTA for 1 week.Decalcified samples were washed with tap water and placed in 70% ethanol. Thehigh levels of radioactivity from Y-90 prevented us from using any instrumentsuch as flow cytometer or fluorescent microscope, and so we had to pursue thefollowing procedures after the radioactivity diminished.
Paraffin-embedded bones were cut into 4-mmslides with bone marrow exposure. Bone marrow morphology was assessed byhematoxylin and eosin (H&E) staining according to standard procedure.
