Thrombopoietin

Manufactured by Thermo Fisher Scientific

Sourced in United States, Canada, United Kingdom

Thrombopoietin is a glycoprotein that regulates the production and differentiation of platelets in the bone marrow. It is a key regulator of megakaryocyte and platelet production.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Mouse thrombopoietin (7 citations) Human thrombopoietin (6 citations) Murine thrombopoietin (3 citations) Murine thrombopoietin (TPO) (3 citations) Human thrombopoietin (TPO) (3 citations) Human thrombopoietin (hTPO) (3 citations) Rh thrombopoietin (2 citations) Recombinant mouse thrombopoietin (mTPO; (2 citations) Rh-thrombopoietin (TPO; (2 citations) Recombinant murine thrombopoietin (2 citations)

68 protocols using thrombopoietin

Placental CD34+ Cells for PNK Generation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Placental CD34⁺ cells were acquired from healthy donors under fully informed consent. With donor eligibility documentation, tissues were qualified using a series of tests including serology and bacteriology (Lifebank USA). Blood was isolated from healthy donor tissues and processed by red blood cell depletion using Hetastarch (Hospira). The resulting cells were then magnetically labeled using Direct CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec). CD34⁺ cells were positively selected using AutoMACS Cell Separator following manufacturer’s protocol. Placental CD34⁺ cells were then cryopreserved in CryoStor CS10 (Biolife Solutions) and stored in liquid nitrogen before use.
For PNK culture, placental CD34⁺ cells were thawed and cultivated following a three-stage process in the presence of cytokines, including thrombopoietin, SCF, Flt3 ligand, IL-7, IL-15 and IL-2 (Thermo Fisher Scientific), for 35 days to generate PNK cells. Nucleofection of CRISPR reagents was performed at day 5-7 of culture. Cell count and passage were performed every 2–3 days and cell expansion was recorded. At the end of the culture, cell phenotype was evaluated by flow cytometry to confirm that the cells expressed typical NK receptors and cytolytic markers.

Guo X., Mahlakõiv T., Ye Q., Somanchi S., He S., Rana H., DiFiglia A., Gleason J., van der Touw W., Hariri R, & Zhang X. (2021). CBLB ablation with CRISPR/Cas9 enhances cytotoxicity of human placental stem cell-derived NK cells for cancer immunotherapy. Journal for Immunotherapy of Cancer, 9(3), e001975.

+ Open protocol

+ Expand

Lentiviral Knockdown of Mtf2 in Mouse Hematopoietic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Adult mouse BM was isolated and lineage-depleted to enrich for stem and progenitor cells (Stem Cell Technologies). Cells were maintained in Iscove's modified Dulbecco's media containing bovine serum albumin, insulin and transferrin (Stem Cell Technologies), 100 U/mL penicillin–streptomycin (ThermoFisher), stem cell factor (SCF; 50 ng/mL), thrombopoietin (10 ng/mL), FLT3 (10 ng/mL) and interleukin-6 (IL-6; 10 ng/mL). Growth factors were purchased from Peprotech. On day 1 of infection, cells were incubated with polybrene (6 mg/mL) for 2 h at 37 °C, and then combined with viral supernatants containing either a green fluorescent protein (GFP)-tagged Mtf2 shRNA clone or a scrambled shRNA control (ThermoFisher). Cells were pelleted at 400 × g for 20 min, and then maintained at 37 °C. On day 2, infection was repeated. Cells were grown for 3 days using a fed-batch culture system and then sorted according to GFP expression. High GFP cells were fixed in 4% PFA, permeabilized with 0.3% Triton and labeled with antibodies against Mtf2 (Genway), Ezh2 (Cell Signaling), Suz12 (Millipore) or H3K27me3 (Millipore) and stained with the appropriate secondary antibodies. Protein expression was determined by flow cytometry compared with an isotype-only control. Data analysis compared mean fluorescent intensity values using a ratio paired t test.

Rothberg J.L., Maganti H.B., Jrade H., Porter C.J., Palidwor G.A., Cafariello C., Battaion H.L., Khan S.T., Perkins T.J., Paulson R.F., Ito C.Y, & Stanford W.L. (2018). Mtf2-PRC2 control of canonical Wnt signaling is required for definitive erythropoiesis. Cell Discovery, 4, 21.

+ Open protocol

+ Expand

Osteogenic Differentiation of Human Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

MSCs and EPCs were purchased from Melero-Martin (Boston, MA, USA) and HSCs from Lonza (Bend, OR, USA). MSCs were expanded in low-glucose Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific, Tarzana, CA, USA) and 1% penicillin–streptomycin (Thermo Fisher Scientific). HSCs were expanded in serum-free hematopoietic progenitor growth medium (Thermo Fisher Scientific) supplemented with 25 ng/mL stem cell factor, 25 ng/mL FMS-like tyrosine 2 ligand, 25 ng/mL thrombopoietin, and 1% penicillin–streptomycin (Thermo Fisher Scientific). The medium used for osteogenic induction was low-glucose DMEM supplemented with 10% FBS, 1% penicillin–streptomycin, 2 mM L-glutamine, 100 nM dexamethasone, 0.2 nM L-ascorbic acid-2-phosphate, and 10 mM β-glycerophosphate. All cells were cultured at 37°C in a 5% CO₂, 95% humidity incubator. The medium was changed every 3 days.
Ethics approval was not required because no animal or human tissue was involved in the study.

Jia Y., Zhang C., Zheng X, & Gao M. (2021). Co-cultivation of progenitor cells enhanced osteogenic gene expression and angiogenesis potential in vitro. The Journal of International Medical Research, 49(4), 03000605211004024.

+ Open protocol

+ Expand

Evaluating AMG 330's Effects on Immature AML Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

To test the effect of AMG 330 on immature AML cell populations, unsorted cells from thawed AML specimens were cultured in IMDM supplemented with 20% FBS and 10 ng/mL each of IL-3, SCF, G-CSF, and GM-CSF containing no AMG 330 or AMG 330 at either 100 pg/mL or 500 pg/mL (because of limited cell numbers available, experiments could only be conducted with 2 different concentrations of AMG 330) together with CellVue Burgundy dye-labeled T-cells at an E:T = 3:1 cell ratio. After 48 hours, CellVue Burgundy dye-negative primary AML cells were separated using a FACSAria flow cytometer (BD Biosciences), and cells were cultured in IMDM with 20% FBS containing the recombinant human cytokines SCF, interleukin-6, FLT3 ligand, and thrombopoietin (all from Invitrogen), StemRegenin-1 (SR1, Cellagen Technology, San Diego, CA), and penicillin-streptomycin [20 (link)]. Cultures were replenished with fresh medium weekly and kept at 37°C in 3% O₂ and 5% CO₂ for 4 weeks. Defined fractions of the cultures were removed after 2 and 4 weeks and subjected to CFC assays. After 10–14 days at 37°C in 3% O₂ and 5% CO₂, colony forming units-granulocyte and/or monocyte (CFU-GM) of at least 30–50 cells were quantified [20 (link)].

Harrington K.H., Gudgeon C.J., Laszlo G.S., Newhall K.J., Sinclair A.M., Frankel S.R., Kischel R., Chen G, & Walter R.B. (2015). The Broad Anti-AML Activity of the CD33/CD3 BiTE Antibody Construct, AMG 330, Is Impacted by Disease Stage and Risk. PLoS ONE, 10(8), e0135945.

+ Open protocol

+ Expand

Generation of Hematopoietic Stem Cells from iPSCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Patient-specific iPSCs were differentiated into hematopoietic stem cells (HSCs) under defined, serum-free and feeder-free conditions as previously reported.^{20 (link)} Briefly, TrypLE (Invitrogen)-dissociated iPSCs were seeded onto 6-well plates that were precoated with 3 mg cm⁻² human plasma fibronectin (Invitrogen) in mTeSR1 supplemented with an inhibitor of Rho-associated kinase (H1152, Sigma). After 1 day, mTeSR1 was replaced with a hematopoietic commitment medium containing Iscove's modified Dulbecco's medium (Invitrogen) supplemented with 1 × HIT (Stem Cell Technologies), 450 μM monothioglycerol (Sigma), 50 ng ml⁻¹ recombinant human BMP4 (R&D Systems), 50 ng ml⁻¹ recombinant human vascular endothelial growth factor (Invitrogen), 0.1 mM NEAA (Invitrogen) and 2 mM L-glutamine (Invitrogen). After 6 days, the medium was changed to a hematopoietic maturation medium consisting of Iscove's modified Dulbecco's medium (Invitrogen) supplemented with 5 U ml⁻¹ heparin (Sigma), 25 ng ml⁻¹ thrombopoietin (Invitrogen), 25 ng ml⁻¹ human recombinant stem cell factor (Invitrogen), 25 ng ml⁻¹ human recombinant FLT3L (Peprotech, Rocky Hill, NJ, USA), 10 ng ml⁻¹ interleukin-3 (Invitrogen) and 10 ng ml⁻¹ interleukin-6 (Invitrogen). The cells were incubated under hypoxic conditions (5% O₂ balanced with nitrogen).

Son M.Y., Lee M.O., Jeon H., Seol B., Kim J.H., Chang J.S, & Cho Y.S. (2016). Generation and characterization of integration-free induced pluripotent stem cells from patients with autoimmune disease. Experimental & Molecular Medicine, 48(5), e232-.

+ Open protocol

+ Expand

Murine Megakaryocyte Isolation and Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the isolation of murine megakaryocytes, bone marrow cells were harvested by flushing the femurs and tibiae with phosphate-buffered saline (Gibco). The obtained cells were separated over Percoll (GE Healthcare) and cultured in specific growth medium (MethoCult®, Stemcell) containing 10% foetal bovine serum (Gibco), 20% IMDM with Glutamax (Gibco) and 50 ng/ml thrombopoietin (Invitrogen). After 7 days differentiation into megakaryocytes was tested by microscopy as well as glyocoprotein Ib (GPIbα) staining and western blotting.

Almilaji A., Yan J., Hosseinzadeh Z., Schmid E., Gawaz M, & Lang F. (2016). Up-Regulation of Na+/Ca2+ Exchange in Megakaryocytes Following TGFβ1 Treatment. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 39(2).

+ Open protocol

+ Expand

Enhancing Vasculogenic Potential of EPCs

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human PBMNCs were isolated as described previously, and 1 × 10³ PBMNCs were added to each well of a 24-well plate (BD Falcon, Bedford, MA, USA) and cultured in serum-free QQc culture medium for 7 d^{17 (link)–19 (link, link)}. Briefly, QQc culture medium is an optimized combination of growth factors and cytokines (20 ng/mL thrombopoietin, 20 ng/mL interleukin (IL) 6, 100 ng/mL stem cell factor (SCF), 100 ng/mL Flt-3 ligand, and 50 ng/mL vascular endothelial growth factor; all from Peprotech, Rocky Hills, NJ, USA) in serum-free stem cell medium (Stemcell Technologies, Vancouver, Canada). QQc culture was performed in this medium for 7 d, which has been shown to dramatically and optimally expand and enhance the vasculogenic potential of EPCs.

Ohtake T., Kobayashi S., Slavin S., Mochida Y., Ishioka K., Moriya H., Hidaka S., Matsuura R., Sumida M., Katagiri D., Noiri E., Okada K., Mizuno H, & Tanaka R. (2018). Human Peripheral Blood Mononuclear Cells Incubated in Vasculogenic Conditioning Medium Dramatically Improve Ischemia/Reperfusion Acute Kidney Injury in Mice. Cell Transplantation, 27(3), 520-530.

+ Open protocol

+ Expand

Culturing Hematopoietic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Kasumi-1, 293T and U937 cells were cultured as previously described.^{14 (link)} G-CSF (granulocyte colony-stimulating factor)-mobilized peripheral blood CD34+ cells were cultured in Iscove's modified Dulbecco's medium (Life Technologies, Karlsruhe, Germany) supplemented with 20% fetal calf serum, 20 ng/ml Flt-3 l, 20 ng/ml GM-CSF, 20 ng/ml stem cell factor (SCF), 20 ng/ml thrombopoietin, 20 ng/ml interleukin (IL)-6, 10 ng/ml IL-3 (all cytokines were obtained Peprotech, Hamburg, Germany), 100 U/ml penicillin/streptomycin and 2 mM L-glutamine. Retroviral transduction and long-term cultivation were performed as previously described.^{5 (link)} IMR-90 cells (ATCC-CCL-186) were cultured as suggested by the supplier.

Wichmann C., Quagliano-Lo Coco I., Yildiz Ö., Chen-Wichmann L., Weber H., Syzonenko T., Döring C., Brendel C., Ponnusamy K., Kinner A., Brandts C., Henschler R, & Grez M. (2014). Activating c-KIT mutations confer oncogenic cooperativity and rescue RUNX1/ETO-induced DNA damage and apoptosis in human primary CD34+ hematopoietic progenitors. Leukemia, 29(2), 279-289.

+ Open protocol

+ Expand

Culturing Leukemia and Primary CD34+ Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human leukemia cell lines MV4-11 and Kasumi-1 were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). MV4-11 and Kasumi-1 cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium supplemented with 10% and 20% fetal bovine serum, respectively. Cell density was kept between 1×10⁵and 1×10⁶ cells/mL. Cells were cultured at 37 ℃ in a humid incubator supplied with 5% CO₂.
Primary CD34⁺ cells were isolated from cord blood with donors’ consent using the commercial isolation kit (STEMCELL Technologies, EasySep^TM Human CD34 Positive Selection Kit II, Canada). CD34⁺ cells were cultured in StemSpan SFEM II (STEMCELL Technologies, Vancouver, BC, Canada) supplemented with FMS-like tyrosine kinase 3 ligand (FLT3L; 100 ng/mL), thrombopoietin (100 ng/mL), and stem cell factor (100 ng/mL; PeproTech, East Windsor, NJ, USA). The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). Informed consent was taken from all the patients. The study was approved by institutional ethics board of Shanghai Tenth People’s Hospital of Tongji University (22KN250).

Cheng Q., Xia J., Wang K., Zhang Y., Chen Y., Zhong Q., Wang X, & Wu Q. (2022). CRISPR/Cas9 ribonucleoprotein (RNP) complex enables higher viability of transfected cells in genome editing of acute myeloid cells. Annals of Translational Medicine, 10(16), 862.

+ Open protocol

+ Expand

Expansion and Differentiation of Hematopoietic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

CB was obtained from the Royal North Shore Hospital and the Australian Cord Blood Bank. Ethical approval for the use of CB was obtained from the Human Research Ethics Committees of the relevant hospitals and the University of New South Wales (approval numbers: HREC 05188, NSCCH 0602-004M, SESIAHS 08/190). Bone marrow mononuclear cells were obtained from Lonza (Mt Waverly Australia). CB processing and isolation of CD34⁺ HSPC and GLYA⁺ cells are described in the Online Supplementary Methods. CD34⁺ HSPC were expanded for 1 week in Isocove modified Dulbecco media (Life Technologies, Carlsbad, CA, USA) with 20% fetal bovine serum (Trace Scientific, Melbourne, Australia), 100 ng/mL stem cell factor (SCF, Amgen, Thousand Oaks, CA, USA), 100 ng/mL thrombopoietin (Peprotech, Rocky Hill, NJ, USA), 100 ng/mL Flt-3 ligand (FLT-3L, Amgen) (STF), 50 mg/mL gentamycin and 200 mM glutamine. The cells were then cultured in cytokine combinations that force expansion and differentiation along specific lineages as described in our previous study (Online Supplementary Table S1).^{14 (link)} Differentiation was assessed by fluorescence activated cell sorting (FACS) analysis after staining cells with the conjugated antibodies detailed in Online Supplementary Table S2. Green fluorescent protein-positive (GFP⁺), GLYA⁺ and CD13⁺ subpopulations were purified by FACS using a FACS Diva (Becton Dickinson).

Richards L.A., Kumari A., Knezevic K., Thoms J.A., von Jonquieres G., Napier C.E., Ali Z., O’Brien R., Marks-Bluth J., Maritz M.F., Pickett H.A., Morris J., Pimanda J.E, & MacKenzie K.L. (2020). DKC1 is a transcriptional target of GATA1 and drives upregulation of telomerase activity in normal human erythroblasts. Haematologica, 105(6), 1517-1526.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!