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Murine thrombopoietin tpo

Manufactured by Thermo Fisher Scientific

Murine thrombopoietin (TPO) is a recombinant protein that functions as a growth factor, specifically stimulating the proliferation and differentiation of megakaryocytes, the precursor cells to platelets. The core function of this product is to support in vitro cell culture research related to platelet production and megakaryocyte biology.

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3 protocols using murine thrombopoietin tpo

1

AML Transformation Assay using Leukemic Cells

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Cells were plated in MethoCult GF M3534 (STEMCELL Technologies, Inc.). Colony numbers were counted and cells were collected, pooled, and replated at the indicated cell numbers per 35‐mm plate. For AE and AE9a colony replating assay, c‐Kit+ bone marrow (BM) cells were isolated by magnetic‐activated cell sorting (MACS) (Miltenyi Biotec) and then cultured in medium containing 100 ng/ml murine stem cell factor (SCF), 100 ng/ml murine thrombopoietin (TPO) and 20 ng/ml murine interleukin‐6 (IL‐6) (Peprotech). The cells were subsequently infected with a retrovirus vector (MigR1‐AE, Addgene plasmid #12431; MigR1‐AE9a, Addgene plasmid #12433, respectively) twice, with a 24‐h interval. Flow cytometry‐sorted EGFP+ cells were used for further experiments. For RNA‐sequencing (RNA‐seq), RNAs were extracted from colony forming cells of the first plating round.
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2

Megakaryocyte Differentiation from Murine Bone Marrow

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Human megakaryoblastic Meg-01 cells were cultured in RPMI 1640 medium at 37°C in 5% CO. Human 293T, hamster BHK-21, and mosquito C6/36 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) at 37°C in 5% CO2, except for C6/36 cells, which were grown at 28°C. All media were supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 50 μg/mL gentamicin. Megakaryocyte differentiation from bone marrow cells of ProT Tg and C57BL/6 mice were maintained in DMEM supplemented with 10% FBS, 50 ng/mL murine thrombopoietin (TPO) (Peprotech, Cranbury, NJ), 10 ng/mL murine IL-3 (Peprotech), 2 mM L-glutamine, and 50 μg/mL gentamicin for 6 days.
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3

Hematopoietic Stem Cell Therapy for X-linked CGD

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BM cells were collected by flushing from femurs and tibias of a pool of male and female XCGD mice. Lin HSPCs isolated using the Lineage Cell Depletion Kit (Miltenyi) were cultured in Stem Span serum-free expansion medium (Stem Cell Technologies) supplemented with 1% l-glutamine, 1% penicillin/streptomycin, murine stem cell factor (SCF; 100 ng/mL), murine thrombopoietin (TPO; 50 ng/mL), human Flt3L (100 ng/mL), and human IL-3 (20 ng/mL; all from PeproTech) for 2 h. Cells were then cultured in the presence or absence (Mock) of a clinical preparation of MSP.Gp91.2T LV at an MOI of 200 for 16 ± 2 h.
Untransduced and transduced cells (1 × 106/mouse) were injected intravenously in lethally (9 Gy split into two equal doses) irradiated 7- to 9-week-old XCGD male mice (XCGD GT mice, n = 20). Only male mice have been used in consideration of the fact that X-linked CGD usually affects only males. Mice were monitored for 52 weeks. Lin HSPCs isolated from XCGD mice and transduced or not with the MSP.Gp91.2T LV (0.5 × 106/mouse) were injected intravenously in lethally (9 Gy split into two equal doses) irradiated 7- to 9-week-old WT CD45.1 male and female mice (XCGD GT WT mice, n = 12), and mice were monitored for 34 weeks.
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