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Isolated bone marrow was separated for different lineages and counted.
On the cross-section the cortical bone and spongy bone was separated manually by auto trace, later the trabeculae and the bone marrow was separated by the threshold function (Fig. 2).
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Mononuclear cells from peripheral blood or bone marrow were separated by Ficoll gradient centrifugation and immunolabeled with PE-anti-Sca-1 (phycoerythrin-conjugated stem cell antigen-1) and APC-anti-Flk-1 [allophycocyanin-conjugated fetal liver kinase-1 (vascularvascular endothelial growth factor receptor 2), 1 µg; BD BioSciences, San Jose, CA].
The bone marrow was centrifuged and cells were fractionated on a density gradient (Biocoll separating solution; Biochrom, Berlin, Germany).
A three-material decomposition (bone, red bone marrow, yellow bone marrow) was performed for each voxel.
Bone marrow was removed with saline lavage.
Bone marrow was obtained from the patients.
ALL diagnostic bone marrow samples were housed in a tumor bank in Children's National Medical Center in Washington, D.C. Mononuclear cells from diagnostic bone marrow aspirates were separated using density centrifugation on Cappel Lymphocyte Separation Medium (ICN Biomedicals, Aurora, Ohio) and immediately flash frozen according to manufacturer's instruction.
Briefly, bone marrow cells were separated using a 3-layered gradient of 1.110 g/ml, 1.090 g/ml, and 1.083 g/ml Percoll PLUS (Amersham, Sweden).
5T2 MM bone marrow cells were separated according to their sedimentation velocity (s.v).v
All the samples of peripheral blood or bone marrow cells were separated using a Ficoll Hypaque gradient, then cell pellets were immediately stored at −80°C.
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