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However, BM expression levels of CD25 and TGFβ1 (Treg-related genes) were equivalent in MM patients and healthy controls, and IL-17 (Th17-related gene) showed no expression in normal or MM bone marrow aspirates using two different primer sequences.
These undifferentiated mesenchymal precursor cells (MPCs) can be isolated from bone marrow aspirates using magnetic activated cell sorting in combination with antibodies that identify STRO-1, VCAM-1 (CD106), STRO-3 (tissue nonspecific alkaline phosphatase), STRO-4 (HSP-90b) and CD146 [ 11, 12].
Genomic DNA was isolated from Jurkat, U937, Bv173, K-562, OC1-AML3, KG-1A and RPMI 8226 cell lines, peripheral blood samples, and bone marrow aspirates using a Blood & Cell Culture DNA Mini Kit from Qiagen GmbH (Hilden, Germany) and following the manufacturer's instructions.
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The thirteen different human NB cell lines or clones, established from 8 patients' tumors or bone marrow aspirates, used for these studies have been published previously (4).
Kinetoplast DNA of L. infantum was detected in the bone marrow aspirate using PCR-ELISA [ 11].
Mononuclear cells were isolated from bone marrow (BM) aspirates using density gradient centrifugation Histopaque-10777; Sigma-Aldrich, St. Louis, MO).
The bone marrow aspirates were used to obtain slide smears that were Giemsa-stained and examined under the optical microscope in order to identify amastigote forms of Leishmania.
Human bone marrow aspirates were used to isolate MSC.
Prior to GEP MMPC need to be enriched resulting in an inability to immediately freeze bone marrow aspirates or use RNA stabilization reagents.
Briefly, marrow cells were aspirated using a 25-gauge needle and a 5 mL syringe containing media.
A composite of synthetic bone substitute (ceraform; calcium phosphate hydroxyapatite 65% and tricalcium phosphate 35%, Teknimed S.A. France) and bone marrow aspirate was used to fill the remaining gaps around the graft.
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