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MSCs from young Fisher-344 rat bone marrow were purified and transfected with pCMV Shh plasmid (ShhMSCs).
Mononuclear cells from bone marrow were purified on a Ficoll gradient at the time of in vitro chemosensitivity assays, frozen in FBS with 10% DMSO, and stored in liquid nitrogen.
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Briefly, CD138+ bone marrow PCs were purified using auto-magnetic-activated cell sorting with anti-CD138 immunobeads as published14.
Bone marrow neutrophils were purified as previously described [27].
Mononuclear bone marrow cells were purified by density gradient centrifugation in Nycoprep 1.077A (Medinor ASA, Norway).
Briefly, hematopoietic bone marrow cells were purified with a 40-μm cell strainer, cultured for 16 hr with 5 ng/ml MCSF (R&DSystems, Minneapolis, Minnesota) in α-MEM containing 10% FBS.
In order to test the age-dependent malignancy of transformed hematopoietic progenitor cells in vivo in the least biased manner possible, and to exclude any potential non cell-autonomous, age-related, effect on the disease evolution, bone marrow cells were purified from CombitTA-BCR-ABLp190 sacriniced mice in which BCR-ABL expression had been kept repressed all life.
The nuclear extracts of bone marrow derived macrophages (5×106) were purified after LPS stimulation.
Peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMC) were purified by Ficoll-Hypaque density gradient centrifugation from 10 40ml PB or BM.
Some of the bone marrow samples and blood samples were purified for the mononuclear cells (including the stem cells in the bone marrow samples).
Recently, Majka et al performed a microarray analysis of bone marrow progenitor-derived adipocytes which were purified from peripheral adipose depots of transplanted recipient mice by flow cytometry [ 20].
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