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Clinimacs cd34 reagent

Manufactured by Miltenyi Biotec
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Sourced in Austria, Germany

The CliniMACS CD34 reagent is a laboratory tool used for the isolation and enrichment of CD34+ cells from human samples. It facilitates the selection and separation of CD34+ cells, which are important for various cell-based therapies and research applications.

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Lab products found in correlation

Lsm 1077 lymphocyte separation medium, by GE Healthcare (1 mentions) Clinimacs instrument, by Miltenyi Biotec (1 mentions) Ack lysing buffer, by Lonza (1 mentions) Heparin, by Merck Group (1 mentions) Human holo transferrin, by Merck Group (1 mentions) Recombinant human insulin, by Merck Group (1 mentions) 7 aad, by Beckman Coulter (1 mentions)

Spelling variants of this product

CliniMACS (62 citations) CliniMACS CD34 Reagent System (17 citations)

4 protocols using clinimacs cd34 reagent

1

Isolation and Enrichment of CD34+ Umbilical Cord Blood Cells

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Mononuclear cells were isolated from the umbilical cord blood samples by density gradient centrifugation (LSM 1077 Lymphocyte Separation Medium, PAA Laboratories GmbH, Graz, Austria) and labelled with CliniMACS CD34 reagent (Miltenyi Biotech, Bergisch-Gladbach, Germany). The selection of CD34+ cells was performed according to the manufacturer's instructions. After the enrichment procedure, the CD34+ cell fraction was collected and cell number and purity were analyzed by flow cytometry. Finally, the obtained CD34+ umbilical cord blood cells were used for the NK cell generation process.
Lehmann D., Spanholtz J., Sturtzel C., Tordoir M., Schlechta B., Groenewegen D, & Hofer E. (2014). IL-12 Directs Further Maturation of Ex Vivo Differentiated NK Cells with Improved Therapeutic Potential. PLoS ONE, 9(1), e87131.
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2

Isolation of CD34+ Cells from Discarded Pediatric Transfusion Blood

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Discarded whole blood from partial manual exchange transfusions was collected from five pediatric research subjects with HbSS genotype (ages 9–16 years) who were at steady state and not being treated with hydroxyurea. The partial manual exchange transfusions were performed as primary (Donors 1–4) or secondary (Donor 5) stroke prophylaxis every 4 to 5 weeks. The chronic transfusion regimens had been in place for a range of 3 to 11 years at the time of this study. Patient's clinical features are described in Table 1. After the RBCs were lysed (ACK Lysing Buffer, Lonza, Walkersville, MD), CD34(+) cells were isolated using CD34 antibodies conjugated to magnetic microbeads (CliniMACS CD34 Reagent, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and a magnetic column (CliniMACS Instrument, Miltenyi Biotec).
de Vasconcellos J.F., Fasano R.M., Lee Y.T., Kaushal M., Byrnes C., Meier E.R., Anderson M., Rabel A., Braylan R., Stroncek D.F, & Miller J.L. (2014). LIN28A Expression Reduces Sickling of Cultured Human Erythrocytes. PLoS ONE, 9(9), e106924.
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3

CRISPR-Mediated Editing of CD34+ HSPCs

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Fresh G-CSF mobilized peripheral blood cells from healthy donor 1 were obtained from Miltenyi Biotec (Auburn, CA). CD34+ HSPCs were isolated using CliniMACS® CD34 reagent (Miltenyi, 130-017-501). Cryopreserved human CD34+ HSPCs from mobilized peripheral blood of deidentified healthy donors 2–7 were obtained from the Fred Hutchinson Cancer Research Center (Seattle, Washington). CD34+ HSPCs were cultured into Stem Cell Growth Medium (SCGM) (CellGenix, 20806–0500) supplemented with 100 ng ml −1 human Stem Cell Growth Factor (SCF) (CellGenix, 1418–050), 100 ng ml −1 human thrombopoietin (TPO) (CellGenix, 1417–050) and 100 ng ml −1 recombinant human FMS-like Tyrosine Kinase 3 Ligand (Flt3-L) (CellGenix cat# 1415–050). HSPCs were electroporated with 3xNLS-SpCas9:sg1617 RNP or HiFi-3xNLS-SpCas9:sg1617 RNP 24 h after thawing. Twenty-four hours after electroporation, HSPCs were transferred into erythroid differentiation medium (EDM) consisting of IMDM (LIFE, 12440061) supplemented with 330 μg ml −1 holo-human transferrin (Sigma, T0665-1G), 10 μg ml −1 recombinant human insulin (Sigma, 19278-5ML), 2 IU ml−1 heparin (Sigma, H3149), 5% human solvent detergent pooled plasma AB (Rhode Island Blood Center), and 3 IU ml −1 erythropoietin (Pharmacy). Five days after electroporation, cells were harvested for gDNA extraction.
Cancellieri S., Zeng J., Lin L.Y., Tognon M., Nguyen M.A., Lin J., Bombieri N., Maitland S.A., Ciuculescu M.F., Katta V., Tsai S.Q., Armant M., Wolfe S.A., Giugno R., Bauer D.E, & Pinello L. (2022). Human genetic diversity alters off-target outcomes of therapeutic gene editing. Nature genetics, 55(1), 34-43.
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4

CD34+ Cell Enrichment Protocol

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The cell suspension was blocked for 5 min at 4 °C with 100 ul/108 cells of Planngamma 100 mg/mL (Griffols). CliniMACS CD34 reagent (Miltenyi) immunomagnetics beads were added at 100 ul/108 cells and incubated for 30 min at room temperature, mixing every 5 min. Subsequently the cells were washed in 50 mL of washing buffer and centrifuged 15 min at 300 g. The labeled cells were then passed through a clinical grade separation mini column (MACS ART MS columns Miltenyi) located in a fixed magnetic field. For each 108 cells two consecutive columns were used. CD45+ and CD34+ content and viability were assessed right after purification and after o/n incubation by flow cytometry using anti-CD45 (clone J33, Beckman), anti-CD34 (clone 581, Beckman) and 7AAD (Beckman) for viability.
Álvarez-Palomo B., Veiga A., Raya A., Codinach M., Torrents S., Ponce Verdugo L., Rodriguez-Aierbe C., Cuellar L., Alenda R., Arbona C., Hernández-Maraver D., Fusté C, & Querol S. (2022). Public Cord Blood Banks as a source of starting material for clinical grade HLA-homozygous induced pluripotent stem cells. Stem Cell Research & Therapy, 13, 408.
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