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Murine tpo

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Sourced in Israel, United States

Murine TPO is a recombinant protein that functions as the thrombopoietin (TPO) cytokine in mice. TPO is the primary regulator of platelet production.

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Lab products found in correlation

Murine scf, by Thermo Fisher Scientific (4 mentions) Doxycycline, by Merck Group (1 mentions) 2 mercaptoethanol, by Merck Group (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Human il 6, by Thermo Fisher Scientific (1 mentions) Human flt3l, by Thermo Fisher Scientific (1 mentions) Recombinant murine il 3, by Thermo Fisher Scientific (1 mentions) Recombinant murine scf, by Thermo Fisher Scientific (1 mentions) Stemspan serum free expansion medium, by STEMCELL (1 mentions) Recombinant human il 6, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Merck Group (1 mentions) L glutamine, by Lonza (1 mentions) Insulin, by Thermo Fisher Scientific (1 mentions) Human holo transferrin, by Merck Group (1 mentions) Mm hepes, by Thermo Fisher Scientific (1 mentions) Ethanolamine, by Merck Group (1 mentions) Polyvinyl alcohol (pva), by Merck Group (1 mentions) Sodium selenite, by Merck Group (1 mentions) Anti biotin magnetic microbeads, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Recombinant murine TPO Murine SCF and TPO Murine thrombopoietin (TPO)

6 protocols using murine tpo

1

Murine Hematopoietic Cell Culture

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Primary cells were cultured in Biotarget-1 serum-free medium supplemented with 2% L-glutamine, 1% penicillin-streptomycin, 1 mM sodium pyruvate, 1% nonessential amino acid solution (all from Biological Industries, Beit Haemek, Israel), 0.06 mM 2-mercaptoethanol (Sigma-Aldrich) and 2 ng/ml doxycycline. The cytokines (PeproTech, Rehovot, Israel) that were added to the growth medium were as follows: murine SCF, murine TPO, murine IL-3 and murine FLT3L (all 10 ng/ml).
Olender L., Bujanover N., Sharabi O., Goldstein O, & Gazit R. (2020). Cyclosporine H Improves the Multi-Vector Lentiviral Transduction of Murine Haematopoietic Progenitors and Stem Cells. Scientific Reports, 10, 1812.
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2

Murine and Human Bone Marrow Cell Isolation and Culture

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Murine bone marrow cells were isolated by flushing femur and tibia of the hind legs of two to three mice. Bone marrow cells from individual animals were pooled for further processing. Mouse bone marrow cells were cultured in DMEM with 10% h.i. FCS (Sigma-Aldrich), 2 mM L-Glutamine (Lonza), 100 U/ml Penicillin and 100 μg/ml Streptomycin (Life Technologies) supplemented with 10ng/ml murine recombinant IL3, 10 ng/ml human recombinant IL6, and 100ng/ml murine recombinant SCF (all Peprotech) (Argiropoulos et al., 2008 (link)).
CD34+ mononuclear cells were isolated from human bone marrow aspirates. Bone marrow aspiration from healthy donors was performed at the Institute of Transfusion Medicine and Immunohematology of Goethe University and German Red Cross Blood Donor Service in Frankfurt. Use of the bone marrow aspirates for research purposes was approved by the Ethics Committee of the University of Frankfurt (329-10) and donors gave written consent for use of the samples. Short term culture of CD34+ bone marrow cells from healthy donors was done in StemSpan™ Serum-Free Expansion Medium (SFEM, Stemcell Technologies) supplemented with 100 ng/ml murine SCF, 100 ng/ml murine TPO, 100 ng/ml human FLT3-L and 100 ng/ml human IL6 (all Peprotech).
Mohr S., Doebele C., Comoglio F., Berg T., Beck J., Bohnenberger H., Alexe G., Corso J., Ströbel P., Wachter A., Beissbarth T., Schnütgen F., Cremer A., Haetscher N., Göllner S., Rouhi A., Palmqvist L., Rieger M.A., Schroeder T., Bönig H., Müller-Tidow C., Kuchenbauer F., Schütz E., Green A.R., Urlaub H., Stegmaier K., Humphries R.K., Serve H, & Oellerich T. (2017). Hoxa9 and Meis1 Cooperatively Induce Addiction to Syk Signaling by Suppressing miR-146a in Acute Myeloid Leukemia. Cancer Cell, 31(4), 549-562.e11.
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3

Albumin-Free Murine HSC Expansion

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HSCs were cultured using a PVA-based albumin-free culture system as
described in a previous report (Wilkinson
et al., 2019). Briefly, HSCs were cultured in media composed of
F12 medium (Gibco), 10 mg/L insulin (Gibco), 6.7 μg/L sodium selenite
(Sigma), 2 mg/L ethanolamine (Sigma), 1% P/S and L-Glutamine (Gibco), 10 mM
HEPES (Gibco), 0.1% poly-vinyl alcohol (PVA, Sigma), 100 ng/ml murine TPO
(PeproTech), 10 ng/ml murine SCF (PeproTech), and human holo-transferrin
(Sigma) with defined concentrations ranging from 5 μg/L to 500 mg/L.
Fifty sorted Lin− Sca-1+ c-Kit+CD48− CD150+ CD34− HSCs
were used as input, and cultured in human fibronectin coated 96-well
microplates (R&D systems) for 18–21 days. Complete medium change
was initiated 5 days after culture started, and performed every 2–3
days. Cultured cells were split 1:3 into new plates at ~90% confluency, and
harvested for analyses and transplantation when they reach confluency for
the second time.
Zhang D., Gao X., Li H., Borger D.K., Wei Q., Yang E., Xu C., Pinho S, & Frenette P.S. (2022). The microbiota regulates hematopoietic stem cell fate decisions by controlling iron availability in bone marrow. Cell stem cell, 29(2), 232-247.e7.
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4

Murine Megakaryocyte Isolation and Culture

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Murine bones (2× femura and 2x tibiae) were removed and bone marrow was flushed in KDS-BSS with 2% FCS. The cells were lineage depleted by incubation with a mix of biotinylated antibodies (CD4, CD2, CD3, CD5, CD8, Ter119, B220, CD19, Gr-1, Ly6G, F4/F8, CD127; WEHI) in KDS-BSS 2% FCS, followed by anti-biotin magnetic microbeads (Miltenyi Biotec, NSW, Australia) and MAC LS columns (Miltenyi Biotec). Single cell suspensions were cultured for 3–5 days at 5 × 105 cells per ml in serum-free medium (StemProTM-34 SFM), supplemented with 100 ng/ml murine TPO (Peprotech) at 37 °C, 5% CO2 and mature megakaryocytes purified using a discontinuous BSA density gradient (3, 1.5 and 0%). Cells were harvested in the 3% layer after 35 min.
Moujalled D., Gangatirkar P., Kauppi M., Corbin J., Lebois M., Murphy J.M., Lalaoui N., Hildebrand J.M., Silke J., Alexander W.S, & Josefsson E.C. (2021). The necroptotic cell death pathway operates in megakaryocytes, but not in platelet synthesis. Cell Death & Disease, 12(1), 133.
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5

Bone Marrow Cell Culture Protocol

Cited 1 time
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BM cultures were performed as described previously (Singh et al., 2013 (link)). In brief, 2 × 105 cells of total nucleated BM cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) medium with 10% serum in presence of the indicated growth factors, 100 ng/ml murine SCF, 50 ng/ml murine IL6, 100 ng/ml murine TPO, and 100 ng/ml murine FLT3L (all cytokines form PeproTech, Rocky Hill, United States) for 10 days.
Liu Y., Chen Y., Deng X, & Zhou J. (2020). ATF3 Prevents Stress-Induced Hematopoietic Stem Cell Exhaustion. Frontiers in Cell and Developmental Biology, 8, 585771.
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6

Purification and Culture of Fetal Mouse HSCs

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The plan for this study was reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Ewha Womans University (No. 16-050). Timed pregnant mice were purchased from a commercial vendor (Orientbio Inc.). Mouse fetal livers were isolated from embryonic day (E) 12.5 and turned into single cell suspension by pipetting and passing through cell strainer. After removing red blood cells, HSCs were purified using The EasySep Mouse Hematopoietic Progenitor Cell Enrichment Kit (STEMCELL Technologies Inc.) following the manufacturer’s protocols. Details are available upon request. Purified cells were seeded in 35 mm dishes (~2 × 105) and cultured for 5 days in StemSpanTM Serum-Free Expansion Medium (STEMCELL Technologies Inc.). Murine TPO (Peprotech) and (R)-TEMOSPho were added at the indicate concentrations. (R)-TEMOSPho was synthesized following previously published steps with minor modifications (14 (link)). Details are available upon request.
Kim J., Jin G., Lee J., Lee K., Bae Y.S, & Kim J. (2019). 2-(trimethylammonium)ethyl (R)-3-methoxy-3-oxo-2-stearamidopropyl phosphate enhances thrombopoietin–induced megakaryocytic differentiation and plateletogenesis. BMB Reports, 52(7), 434-438.
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