NaHS (Sigma-Aldrich, St. Louis, MO, USA) was applied as a donor of H2S [45 (link)] in the present study. For the in vivo administration, NaHS and recombinant murine TPO (R&D, NASDAQ, USA) were freshly diluted in sterile saline or PBS for each administration. A single dose of TPO (15 μg kg−1) was given by intraperitoneal injection after radiation exposure whereas NaHS was injected once a day during the experimental period. For in vitro experiments, recombinant murine TPO (PeproTech, NJ, USA) was freshly diluted with cell culture medium and added to the cultured cells with a single dose of 10 ng mL−1 given at the first day of the experiments. NaHS was diluted with cell culture medium and added to the cultured cells every 8 h.
Recombinant murine tpo
Manufactured by Thermo Fisher Scientific
Sourced in United States
Recombinant murine TPO is a protein that functions as the primary regulator of megakaryocyte production and platelet formation.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant murine scf, by Thermo Fisher Scientific (3 mentions)
Recombinant murine il 12, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fortessa 2, by BD (1 mentions)
Recombinant murine il 7, by Thermo Fisher Scientific (1 mentions)
Recombinant murine il 3, by Thermo Fisher Scientific (1 mentions)
Recombinant murine il 6, by Thermo Fisher Scientific (1 mentions)
Recombinant murine il 5, by Thermo Fisher Scientific (1 mentions)
Recombinant murine il 11, by Thermo Fisher Scientific (1 mentions)
Recombinant murine il 4, by Thermo Fisher Scientific (1 mentions)
Recombinant murine gm csf, by Thermo Fisher Scientific (1 mentions)
4 protocols using recombinant murine tpo
1
Hydrogen Sulfide and Thrombopoietin in Radiation Exposure
2
Single-Cell Hematopoietic Stem Cell Expansion
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Briefly, 60 HSC1 cells were individually sorted into a 96-well U-bottom plates, with each well receiving 200 μL of the new standard serum-free medium supplemented with 50 ng/mL recombinant murine SCF (Peprotech, 250-03), 50 ng/mL recombinant murine TPO (Peprotech, 315-14), and 50 ng/mL recombinant murine IL-12 (Peprotech, Rocky Hill, NJ, 210-12). Single cells were cultured at 37°C in a humidified atmosphere with 5% CO2 for 7 days. The cell numbers in each well was visually counted every day using an inverted microscope. The first division kinetics was obtained based on the cumulative number of wells where ≥2 cells were found during 7 days of culture. The second-division kinetics were obtained based on the cumulative number of wells where ≥3 cells were found. The third-division kinetics were obtained based on the cumulative number of wells where ≥5 cells were found.
3
Single-cell CFSE+ HSC Culture
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Single-cell–sorted CFSE+ HSC (see “CFSE cell staining dye” section) were seeded onto mitomycin C–treated OP9 feeder cells. Cell was plated in one well of a 96-well plate and grown in 200 μl of IMDM-10% medium at 37°C and 5% CO2. For maintenance the sorted HSCs were kept in IMDM medium (Gibco, #12440053), supplemented with 1x penicillin/streptomycin (Gibco, 10378016), 10% FCS, recombinant murine SCF (100 ng/ml; PeproTech, #250-03), and recombinant murine TPO (10 ng/ml; PeproTech, #315-14).
4
Clonal analysis of mouse hematopoietic stem cells
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Freshly collected mouse BM was labeled according to panels exemplified in Supplementary Data. Single HSCs (LSK+CD34−Flt3−CD150+CD48−) were sorted into 96-well plates, using an Aria FACS Fusion II with a 100-μm nozzle. Cells were cultured for 10 days in IMDM-10% (see the “Cell culture” section), supplemented with SCF (50 ng/ml; recombinant murine SCF, PeproTech, #250-03), interleukin-3 (IL-3; 10 ng/ml; recombinant murine IL-3, PeproTech, #213-13), IL-6 (10 ng/ml; recombinant murine IL-6, PeproTech, #216-16), erythropoietin (EPO; 2 U/ml; BioLegend, #587602), IL-11 (50 ng/ml; recombinant murine IL-11, PeproTech, #220-11), IL-5 (10 ng/ml; recombinant murine IL-5, PeproTech, #215-15), thrombopoietin (TPO; 50 ng/ml; recombinant murine TPO, PeproTech, #315-14), IL-4 (10 ng/ml; recombinant murine IL-4, PeproTech, #214-14), granulocyte-macrophage colony-stimulating factor (GM-CSF; 15 ng/ml; recombinant murine GM-CSF, PeproTech, #315-03), and IL-7 (10 ng/ml; recombinant murine IL-7, PeproTech, #217-17). Growth factors/cytokines were refreshed at day 5. The clones/colonies in each well were labeled on day 10 with anti-CD117, anti-Ter119, anti-CD11b, and anti-IgD antibody. Clones were analyzed by flow cytometry using the BD Fortessa II (BD Biosciences) and processed with FlowJo 10.v.
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