Rs Both hind paws were excised, and a hind leg was immediately frozen YM155 in liquid nitrogen and stored at 0 for future studies of Luciferaseaktivit t. The other posterior branch in 10% formalin and immersed overnight in 70% ethanol until decalcification, and histomorphometric study. The load of bone metastasis of tumors was directly by measuring the luciferase values in hind leg and indirectly by measuring the bone volume percent in the other rear extremity T with Reinholz et al. Page 4 bones. Author manuscript, increases available in PMC 2011 1 July. NIH PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript bone histomorphometry analysis with H Matoxylin and eosin F Staining of tissue sections using the system described OsteoMeasure Fnd Rbt.
E7080 The detection of tumor cells expressing luciferase in lysates bone when testing luciferase, frozen bones were crushed and homogenized in lysis buffer 0.250 ml reporter. Luciferase activity t was in the supernatant of freeze-drying whole homogenates thawed using a luminometer TD20/20 and luciferase assay kit according to manufacturer’s instructions measured. Luciferase activity was t normalized to the total tissue protein using the DC protein assay. Since the measurement of luciferase in bone provides a more comprehensive and less subjective in tumor burden for this test was to compare intramedullary Ren tumor burden. Bone histomorphometry in the distal femoral bone volume percent of a rear leg was determined by bone histomorphometry analysis.
The tissue sections were removed Herk Mmlichen methods and found Rbt with H & E. made Repr Sentative portions of the central portion of each femur were used to determine the volume percent of the bone under a microscope at a magnifying your TION × to determine the 10th Bone volume of the distal femur end-stage mouse was in the L Longitudinal section H & E Fnd Rbten measured using the system OsteoMeasure. Bone volume was measured at 350 m from the growth plate in the two fields of 700 m2 in the same tissue section and the results are expressed as percent bone volume measured by the whole bottle Surface. In vitro tests of multiple myeloma cell proliferation of myeloma cells were obtained from three patients and myeloma cell lines were generated and various referred to as KAS 6/1, 6 DP and KP 6th In vitro cell proliferation was measured by tests of tritiated thymidine by cells as described above.
KAS 1.6 mouse model MIP1 We used an animal model of multiple myeloma in which the systemic administration of CIS 6.1 MIP1 myeloma cells in immunodeficient female M Mice produced progressive bone loss measured by diffuse DMO. In this animal model, the initiation of bone loss occurs within two weeks after tumor cell injection step L Hmungen the hind legs in 7-8 weeks after injection, and death usually occurs within 8-9 weeks after injection. KAS man 1.6 myeloma cells were genetically Changed to the osteoclast-activating factor, MIP1, the loss of bone density from these M Carry nozzles induces observed. No loss of BMD was in SCID-M Mice observed with mothers KAS 6/1 is injected and the Erh Increase in bone density was similar, not the animals injected with tumor cells. However, a allm Hliche decrease in bone density in the femur and lumbar vertebra Column from animals that were injected with t observed