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Macs cd34 microbeads

Manufactured by Miltenyi Biotec
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MACS CD34 MicroBeads are magnetic bead-conjugated antibodies specifically designed for the isolation and enrichment of CD34-positive cells from various sample types. The core function of these microbeads is to enable the separation and purification of CD34-expressing cells using magnetic separation techniques.

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Lab products found in correlation

Pharm lyse, by BD (2 mentions) Collagenase dispase, by Roche (2 mentions) Quadromacs separator, by Miltenyi Biotec (2 mentions) Ack lysing buffer, by Thermo Fisher Scientific (2 mentions) Flt3l, by Thermo Fisher Scientific (1 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Stemspan sfem 2, by STEMCELL (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Pd0332991, by Pfizer (1 mentions) Cytarabine ara c, by Merck Group (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Hl 60, by American Type Culture Collection (1 mentions) Doxycycline, by Takara Bio (1 mentions) Stemdiff hematopoietic kit, by STEMCELL (1 mentions) Gentle cell dissociation reagent (gcdr), by STEMCELL (1 mentions) Matrigel hesc qualified matrix, by Corning (1 mentions)

7 protocols using macs cd34 microbeads

1

Human CD34+ HSC Engraftment in NSG Mice

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Human CD34+ hematopoietic stem cells (HSC) were isolated from fetal livers obtained from Advanced Bioscience Resources, INC (ABR, Alameda, CA). Tissue was disrupted and incubated with 1mg/mL Collagenase/Dispase (Roche Applied Sciences) for 15 min at 37°C. Cells were isolated by passing the disrupted tissue through a 70 μm filter. Red blood cells were lysed in BD Pharm Lyse (BD Biosciences, San Jose, CA), with CD34+ cells being isolated using CD34 MACS microbeads (Miltenyi) according to manufacturer’s instructions with an additional purification step using a second column. NOD.Cg-Prkdc scid Il2rγ tm1Wj/Szj (NOD/SCID/IL2rγnull, NSG) mice were obtained from Jackson Laboratories (Bar Harbor, ME). Neonatal mice received 150 cGy radiation, and 2–4 hours later 1×106 CD34+ HSCs in 1% heparin (Celgene, Summit, NJ) via intrahepatic injection. Mice were monitored for engraftment levels of human CD45+ cells and development of T cells and B cells at 8, 10, and 12 weeks post engraftment.
Gardner M.R., Kattenhorn L.M., Kondur H.R., von Schaewen M., Dorfman T., Chiang J.J., Haworth K.G., Decker J.M., Alpert M.D., Bailey C.C., Neale ES J.r., Fellinger C.H., Joshi V.R., Fuchs S.P., Martinez-Navio J.M., Quinlan B.D., Yao A.Y., Mouquet H., Gorman J., Zhang B., Poignard P., Nussenzweig M.C., Burton D.R., Kwong P.D., Piatak M J.r., Lifson J.D., Gao G., Desrosiers R.C., Evans D.T., Hahn B.H., Ploss A., Cannon P.M., Seaman M.S, & Farzan M. (2015). AAV-expressed eCD4-Ig provides durable protection from multiple SHIV challenges. Nature, 519(7541), 87-91.
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2

Human CD34+ HSC Engraftment in NSG Mice

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Human CD34+ hematopoietic stem cells (HSC) were isolated from fetal livers obtained from Advanced Bioscience Resources, INC (ABR, Alameda, CA). Tissue was disrupted and incubated with 1mg/mL Collagenase/Dispase (Roche Applied Sciences) for 15 min at 37°C. Cells were isolated by passing the disrupted tissue through a 70 μm filter. Red blood cells were lysed in BD Pharm Lyse (BD Biosciences, San Jose, CA), with CD34+ cells being isolated using CD34 MACS microbeads (Miltenyi) according to manufacturer’s instructions with an additional purification step using a second column. NOD.Cg-Prkdc scid Il2rγ tm1Wj/Szj (NOD/SCID/IL2rγnull, NSG) mice were obtained from Jackson Laboratories (Bar Harbor, ME). Neonatal mice received 150 cGy radiation, and 2–4 hours later 1×106 CD34+ HSCs in 1% heparin (Celgene, Summit, NJ) via intrahepatic injection. Mice were monitored for engraftment levels of human CD45+ cells and development of T cells and B cells at 8, 10, and 12 weeks post engraftment.
Gardner M.R., Kattenhorn L.M., Kondur H.R., von Schaewen M., Dorfman T., Chiang J.J., Haworth K.G., Decker J.M., Alpert M.D., Bailey C.C., Neale ES J.r., Fellinger C.H., Joshi V.R., Fuchs S.P., Martinez-Navio J.M., Quinlan B.D., Yao A.Y., Mouquet H., Gorman J., Zhang B., Poignard P., Nussenzweig M.C., Burton D.R., Kwong P.D., Piatak M J.r., Lifson J.D., Gao G., Desrosiers R.C., Evans D.T., Hahn B.H., Ploss A., Cannon P.M., Seaman M.S, & Farzan M. (2015). AAV-expressed eCD4-Ig provides durable protection from multiple SHIV challenges. Nature, 519(7541), 87-91.
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3

Isolation of CD34+ Cells from Umbilical Cord Blood

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We collected samples from consenting donors at the gynecology and obstetrics department of Liaocheng People’s Hospital. Mononuclear cells (MNC) from umbilical cord blood were isolated by density gradient centrifugation using Ficoll-Lympholyte separation (Cedarlane, Burlington, Canada). After labeling of CD34+ cells by MACS CD34 MicroBeads (Miltenyi Biotec, Auburn, CA, USA), CD34 microbead-labeled cells were isolated using an LS column. We performed as the manufacturer’s protocol to operate QuadroMACS separator (Miltenyi Biotec). The purity of the isolated CD34+ cells was about 86.37%±3.56% (n=6). This study was approved by the Ethics Committee of Liaocheng People’s Hospital and Clinical School of Shandong First Medical University (No. 2012004), and was conducted in accordance with the Declaration of Helsinki (as revised in 2013). Written informed consent was obtained from all participants included in this study.
Hua J., Jiao T., Qiao Y., Zhang S., Xiao T., Zhang Y, & Yan J. (2023). Human serum albumin promotes self-renewal and expansion of umbilical cord blood CD34+ hematopoietic stem/progenitor cells. Annals of Translational Medicine, 11(2), 62.
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4

Isolation and Cryopreservation of CB and AML HSPCs

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Mononuclear cells from CB and AML samples were isolated by Ficoll-Paque PLUS (GE Healthcare) density gradient centrifugation, and red blood cells were removed by ACK lysis (ACK Lysing Buffer, Thermo Fisher Scientific). CD34+ HSPCs were enriched from CB mononuclear cells by magnetic cell separation using MACS CD34 MicroBeads (Miltenyi Biotec). AML samples were resuspended in 90% FBS + 10% DMSO (MilliporeSigma) and cryopreserved in liquid nitrogen for future use.
CD34-enriched cells were either cryopreserved for future use or cultured in low-density conditions (200,000 cells/mL) at 37°C in 5% CO2 in StemSpan SFEM II (STEMCELL Technologies) base media supplemented with 20 ng/mL TPO, SCF, FLT3L, and IL-6 (Peprotech), and 35 nM small-molecule UM-171.
Fan A.C., Nakauchi Y., Bai L., Azizi A., Nuno K.A., Zhao F., Köhnke T., Karigane D., Cruz-Hernandez D., Reinisch A., Khatri P, & Majeti R. (2023). RUNX1 loss renders hematopoietic and leukemic cells dependent on IL-3 and sensitive to JAK inhibition. The Journal of Clinical Investigation, 133(19), e167053.
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5

Isolation of Human Umbilical Cord CD34+ Cells

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Samples were collected from consenting donors by Shandong Qilu Stem Cell Engineering Co., Ltd. in accordance with the declaration of Helsinki and approved by the Ethics Review Board of the Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences. Fresh collected human umbilical cord blood CD34+ cells were isolated using LS Column and QuadroMACS Separator (Miltenyi Biotec), according to the manufacturer’s protocol after collecting CD34 MicroBead-labeled cells by magnetic activated cells sorting (MACS) CD34 MicroBeads (Miltenyi Biotec).
He M., Xu H., Liu G., Yang M., Zhang W., Li Y., Zhang H., Wang C., Zhang Y., Liu X., Xu S., Ding Y., Li Y., Gao Y, & Zhang Q. (2022). Levistilide A Promotes Expansion of Human Umbilical Cord Blood Hematopoietic Stem Cells by Enhancing Antioxidant Activity. Frontiers in Pharmacology, 13, 806837.
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6

Isolation and Culture of AML Cell Lines

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The human AML cell line HL-60 and the U937 histiocytic lymphoma with monocytic features were obtained from the American Type Culture Collection (Manassas, VA). The cell lines were maintained in RPMI-1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum, 100 units/mL penicillin, and 100 units/mL streptomycin. The human primary CD34+ AML cells were purified using MACS CD34 MicroBeads (Miltenyl Biotec), and co-cultured with mitomycin-treated HS-5 human stromal cells (17 (link)). The peripheral blood mononuclear cells were isolated using ficoll density centrifugation (GE lifetechology). Cytarabine (Ara-C) was purchased from Sigma. PD 0332991 was obtained from Pfizer (La Jolla, CA).
Yang C., Boyson C.A., Di Liberto M., Huang X., Hannah J., Dorn D.C., Moore M.A., Chen-Kiang S, & Zhou P. (2015). CDK4/6 inhibitor PD 0332991 sensitizes acute myeloid leukemia to cytarabine-mediated cytotoxicity. Cancer research, 75(9), 1838-1845.
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7

Differentiation of BiPSCs into HPCs

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Differentiation of BiPSCs into HPC was performed using STEMdiff Hematopoietic Kit (STEMCELL Technologies, Vancouver, Canada). BiPSCs were cultured on Matrigel hESC-Qualified Matrix (CORNING, Corning, NY, USA), incubated with Gentle Cell Dissociation Reagent (STEMCELL Technologies) at room temperature for 4–6 min, removed using a scraper, and then, 50 μm-sized pieces were transferred to a matrigel-coated 12-well plate in Complete StemFit AK02N (100 pieces/well). Differentiation experiments were performed according to the instructions of the kit. BiPSCs with AID were cultured in the presence of 10 ng/mL doxycycline (Takara Bio, Kusatsu, Japan) to prevent AID expression. Concurrently, CD34+ cells were purified using MACS CD34 MicroBeads (Miltenyi Biotec Inc., Auburn, CA, USA) according to the manufacturer’s instructions, and the collected cells were used for phenotype analysis and a colony-forming assay.
Azami Y., Tsuyama N., Abe Y., Sugai-Takahashi M., Kudo K.I., Ota A., Sivasundaram K., Muramatsu M., Shigemura T., Sasatani M., Hashimoto Y., Saji S., Kamiya K., Hanamura I., Ikezoe T., Onodera M, & Sakai A. (2021). Chromosomal translocation t(11;14) and p53 deletion induced by the CRISPR/Cas9 system in normal B cell-derived iPS cells. Scientific Reports, 11, 5216.
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