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Interleukin 3 il 3

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Interleukin-3 (IL-3) is a cytokine that stimulates the growth and differentiation of hematopoietic precursor cells. It promotes the survival, proliferation, and differentiation of multipotent progenitor cells, as well as the maturation of various blood cell types, including granulocytes, erythrocytes, macrophages, and megakaryocytes.

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Lab products found in correlation

Heparin, by Merck Group (1 mentions) M3236, by STEMCELL (1 mentions) Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Stabilized glutamine, by Harvard Bioscience (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Macrophage colony stimulating factor m csf, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) X vivotm 15, by Lonza (1 mentions) Aggrewell, by STEMCELL (1 mentions) β mercaptoethanol, by Thermo Fisher Scientific (1 mentions) Tryple express, by Thermo Fisher Scientific (1 mentions)

4 protocols using interleukin 3 il 3

1

Quantification of Hematopoietic Progenitor Colonies

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After isolation of c-kit+/Sca-1−/Lin− (KSL) cells from mouse BM (Kwon et al., 2011 (link)), the frequencies of small or large colonies was determined after culturing KSL cells (500 cells/35 mm dish) for 10 days in methylcellulose-containing medium M3236 (Stem Cell Technologies) supplemented with 20 ng/ml stem cell factor (SCF), 50 ng/ml VEGF, 20 ng/ml interleukin-3 (IL-3) (R&D Systems), 50 ng/ml bFGF, 50 ng/ml EGF (PeproTech), 2 U/ml heparin (Sigma), 30% FBS, and antibiotics. Colony forming units (CFUs) were counted under an inverted microscope at 40x. In our previous study, we defined small- and large-CFUs for the expression of additional endothelial marker genes such as CD31, Flk-1, von Willebrand factor (vWF), VE-cadherin and eNOS (Kwon et al., 2011 (link)).
Choi J.H., Nguyen M.P., Lee D., Oh G.T, & Lee Y.M. (2014). Hypoxia-Induced Endothelial Progenitor Cell Function Is Blunted in Angiotensinogen Knockout Mice. Molecules and Cells, 37(6), 487-496.
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2

Erythroid Differentiation from Mononuclear Cells

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Cultures were initiated from frozen or freshly isolated mononuclear cells or CD34+ cells. For depletion of leukocytes, mononuclear cells were pre-cultured for 2 days in IMDM with stabilized glutamine (Biochrom, Berlin, Germany) containing 5% (vol/vol) heparinized human plasma and 3 U/ml EPO (Janssen-Cilag, Dublin, Ireland). Cells were then washed and cultured in erythroid proliferation medium [12] (link) consisting of IMDM with 330 μg/ml iron-saturated human transferrin and 10 μg/ml recombinant human insulin, supplemented with 5% heparinized human plasma, 100 ng/ml stem cell factor (SCF) (Cambridge Biosciences, Cambridge, UK), 5 ng/ml interleukin-3 (IL-3) (R&D Systems, Minneapolis, MN, USA), 3 U/ml EPO and 10−6 M hydrocortisone for 1–2 days. CD34+ cells were plated directly into the appropriate conditions. Manual cell counting was performed using the trypan blue exclusion method with trypan blue diluted 1/6 in phosphate buffered saline (PBS) and added to cells at 1:1 or 1:4 ratios.
Boehm D, & Bell A. (2014). Simply red: A novel spectrophotometric erythroid proliferation assay as a tool for erythropoiesis and erythrotoxicity studies. Biotechnology Reports, 4, 34-41.
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3

Differentiation of Human ESCs to Myeloid Cells

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Human ESCs were first differentiated to EBs by dissociation with TrypLE Express (Invitrogen). A total of 300 EBs were generated from ∼4 × 106 cells, aggregated in mTeSR1 medium with 10 mM ROCK inhibitor Y-27632 (Calbiochem) by spinning in an AggreWell (Stem Cell Technologies) according to the manufacturer's manual. The EBs were analyzed at day 4 post-aggregation. For subsequent myeloid differentiation, EBs were cultured in medium consisting of X-VIVOTM15 (Lonza), supplemented with 100 ng/ml Macrophage colony-stimulating factor (M-CSF) (Invitrogen), 25 ng/ml Interleukin 3 (IL-3) (R&D), 62 mM Glutamax (Invitrogen), 100 U/ml Penicillin and 100 μg/ml Streptomycin (Invitrogen) and 0.055 mM β-Mercaptoethanol (Invitrogen). Once ESC-derived monocytes were visible in the supernatant of the cultures (from 2–3 weeks onward), the non-adherent monocytes were harvested weekly, as has been described previously (36 (link)). Monocytes released from the ‘factories’ were used directly.
Vazquez-Arango P., Vowles J., Browne C., Hartfield E., Fernandes H.J., Mandefro B., Sareen D., James W., Wade-Martins R., Cowley S.A., Murphy S, & O'Reilly D. (2016). Variant U1 snRNAs are implicated in human pluripotent stem cell maintenance and neuromuscular disease. Nucleic Acids Research, 44(22), 10960-10973.
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Conditional Deletion of ZBP-89 in Hematopoietic Mice

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Mice expressing the targeted ZBP-89 locus with flanking LoxP sites (ZBP-89fl/fl) were produced as described 11 (link). Conditional deletion of ZBP-89 in the hematopoietic system of mice (C57BL/6-CD45.2+ background) was generated as shown in Figure 1A. Transgenic mice expressing an interferon-inducible Cre recombinase (MxCre) (Kuhn et al, 1995) were kindly provided by Dr. Hanno Hock (MGH Cancer Center). Murine erythroleukemia cell line MEL and the non-hematopoietic QM7 cell line, which lacks endogenous ZBP-89 12 (link), were obtained from ATCC and maintained in DMEM medium containing 10% fetal calf serum. The multipotent A4 cell line was maintained in Iscove’s modified Dulbecco’s Medium (IMDM) supplemented with 20% horse serum and Interleukin-3 (IL-3)(100 units/ml, R&D Systems). Congenic C57BL/6-CD45.1 mice (6 to10 weeks old) were used as donors for purification of wild type cells and as recipients for BM transplantation (BMT) experiments. All animal experiments were performed in accordance with legal and ethical requirements demanded by law and approved by the Massachusetts General Hospital Subcommittee on Research Animal Care.
Li X., Romain R.D., Park D., Scadden D.T., Merchant J.L, & Arnaout M.A. (2014). Stress hematopoiesis is regulated by the Krüppel-like transcription factor ZBP-89. Stem cells (Dayton, Ohio), 32(3), 791-801.
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