Mononuclear cells were isolated from CB (obtained from the Cleveland Blood Center on a contractual basis) by Ficoll-Hypaque Plus density centrifugation. CD34+ hematopoietic stem and progenitor cells (HSPCs) were purified by positive selection using the Midi-magnetic–activated cell sorting LS columns (130-042-401, Miltenyi Biotec). CD34+ cells were cultured in Iscove modified Dulbecco medium (IMDM) containing 20% BIT 9500 medium (Stem Cell Technologies) supplemented with SCF (100 ng/mL), Fms-like tyrosine kinase 3 (FLT-3; 10 ng/mL), IL-6 (20 ng/mL), and thrombopoietin (TPO; 100 ng/mL; these cytokines were purchased from PeproTech).
Midi magnetic
Manufactured by Miltenyi Biotec
Sourced in United States
The Midi-magnetic is a laboratory equipment product designed for the separation and isolation of target cells or molecules from complex biological samples. It utilizes magnetic separation technology to enable efficient and gentle processing of samples. The core function of the Midi-magnetic is to facilitate the selective isolation of desired components from a heterogeneous mixture.
Automatically generated - may contain errors
Lab products found in correlation
Bit 9500 medium, by STEMCELL (1 mentions)
Thrombopoietin tpo, by Thermo Fisher Scientific (1 mentions)
Fms like tyrosine kinase 3 flt3, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Granulocyte macrophage colony stimulating factor gm csf, by Thermo Fisher Scientific (1 mentions)
Iscove s modified dulbecco s medium imdm, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Anti fitc microbeads, by Miltenyi Biotec (1 mentions)
2 protocols using midi magnetic
1
Isolating and Culturing CD34+ Hematopoietic Stem Cells
2
Purification and Culture of ScaI+ Cells
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ScaI + bone marrow (BM) cells were selected using ScaI MultiSort microbeads and midi-magnetic-activated cell sorting (MACS) separation columns (Miltenyi Biotec, Auburn, CA, USA). ScaI + cells were lineage-depleted by labelling with fluorescein isothiocyanate (FITC)-conjugated anti-CD4, anti-CD8, anti-CD11b, anti-Gr-1 and anti-B220 (BD PharMingen), binding to anti-FITC microbeads (Miltenyi Biotec) and passing them through miniMACS separation columns (purity, 95 ± 2% Lin -ScaI + ). Lin -ScaI + cells were grown as suspension cultures for 5 days in Teflon jars (37°C, CO 2 ) with Iscove's modified Dulbecco's medium (IMDM) (Life Technologies, Grand Island, NY, USA), supplemented with 10% FBS (HyClone Laboratories, Logan, UT, USA), 1 ng/ml granulocyte-macrophage colony-stimulating factor (GM-CSF) (Peprotech) and 50 ng/ml p19, Ebi3 or IL-39.
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