BM samples were collected for routine diagnostic or evaluation purposes after patients' informed consent and in accordance with the Declaration of Helsinki. All research was approved by the local ethical committee (B.U.N. 143201316382). BM mononuclear cells were obtained after Ficoll density gradient centrifugation (Nycomed, Lucron Bioproducts, Zurich, Switserland) and purified using MACS Cell Separation MS Columns (Miltenyi Biotec, Bergisch-Gladbach, Germany) and CD138 microbeads according to the manufacturer's protocol. Purity of the samples was evaluated by flow cytometry (FACS Canto, BD Biosciences, Franklin Lakes, USA) and analysed using FACSDiva software (BD Biosciences).
Macs cell separation ms columns
Manufactured by Miltenyi Biotec
Sourced in Germany
The MACS Cell Separation MS Columns are lab equipment used for the magnetic separation of cells. They are designed to work with the MACS technology to isolate target cells from complex sample mixtures.
Automatically generated - may contain errors
Lab products found in correlation
Ficoll density gradient centrifugation, by Takeda Pharmaceuticals (1 mentions)
Facsdiva software, by BD (1 mentions)
Facscanto, by BD (1 mentions)
Cd138 microbeads, by Miltenyi Biotec (1 mentions)
Recombinant il 2, by Thermo Fisher Scientific (1 mentions)
αcd28 37.51, by BioXCell (1 mentions)
αcd3 2c11, by BioXCell (1 mentions)
Cd4 microbeads, by Miltenyi Biotec (1 mentions)
2 protocols using macs cell separation ms columns
1
Isolation of Bone Marrow Mononuclear Cells
2
T Cell Isolation and Activation
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VAT was pooled from two or three mice of the same genotype to ensure adequate cell number for culture. Cells were isolated as described above, and CD4+ T cells were enriched by incubating cells with CD4 MicroBeads and positively selecting with MACS cell separation MS columns (Miltenyi Biotec). Purified cells were resuspended in RPMI-C with 500 U/ml recombinant IL-2 (eBioscience). Cells were cultured for 48 h in 96-well flat-bottom plates with plate-bound αCD3 (2C11) and αCD28 (37.51) from Bio X Cell at 1 µg/µl, with or without the addition of plate-bound αICOS (C398.4A; BioLegend) at 2 µg/µl. Expression of CCR3 was assessed by flow cytometry as described above.
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