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Human tpo

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Human TPO is a laboratory equipment product used for the detection and quantification of human thyroperoxidase (TPO) in biological samples. TPO is an enzyme involved in the production of thyroid hormones. This product allows for the analysis of TPO levels, which can provide insights into thyroid function and associated medical conditions.

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Lab products found in correlation

Human scf, by Thermo Fisher Scientific (7 mentions) Human il 3, by Thermo Fisher Scientific (5 mentions) Human flt3 ligand, by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (2 mentions) Cd34 apc, by BioLegend (2 mentions) Cd38 pe, by BD (2 mentions) Apc cy7 cd49f, by BioLegend (2 mentions) Cd45 pe cy7, by BioLegend (2 mentions) V450 hcd45 antibodies, by BD (2 mentions) Cd90 fitc, by BioLegend (2 mentions) Stemspan sfem, by STEMCELL (2 mentions) Horse serum, by Thermo Fisher Scientific (2 mentions) Mil 3, by Thermo Fisher Scientific (2 mentions) Human flt3l, by Thermo Fisher Scientific (2 mentions) Hil 6, by Thermo Fisher Scientific (2 mentions) Imdm medium, by Thermo Fisher Scientific (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Cd34 microbead kit, by Miltenyi Biotec (2 mentions) Murine il 3, by Thermo Fisher Scientific (2 mentions)

16 protocols using human tpo

1

Expansion and Xenotransplantation of Human Cord Blood CD34+ Cells

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Human umbilical cord blood-derived CD34+ cells (purchased from Lonza or Stem Cell Technologies) or FACS-purified CD34+CD38-CD90+CD49f+ cells (following staining with APC-CD34 [Biolegend; 560940], PE-CD38 [BD; 347687], FITC-CD90 [Biolegend; 328113], and APC/Cy7-CD49f [Biolegend; 313611]) were cultured in IMDM (Life Technologies) containing 0.1% HSA or 0.1% PVA (Sigma P8136, 363081, or 363146), supplemented with 1% ITSX, 1% P/S/G, 10 mM HEPES. For proliferation assays, 50 cells were seeded per well and supplemented with 10 ng/ml human SCF and 100 ng/ml human TPO (Peprotech). During the cultures, media was refreshed every three days and counted at day-7. For xenograft assays, 2×103 cells were expanded for 7-days before being injected intravenously into sub-lethally-irradiated (1.5 Gy) NOG mice. Human cell chimerism in the PB was calculated at 16-weeks post-transplantation using PE/Cy7-CD45.1 (Biolegend; 110730) and V450-hCD45 antibodies (BD; 560367).
Wilkinson A.C., Ishida R., Kikuchi M., Sudo K., Morita M., Crisostomo R.V., Yamamoto R., Loh K.M., Nakamura Y., Watanabe M., Nakauchi H, & Yamazaki S. (2019). Long-term ex vivo hematopoietic stem cell expansion affords nonconditioned transplantation. Nature, 571(7763), 117-121.
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2

CRISPR Editing of Umbilical Cord HSCs

Cited 1 time
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Umbilical cord blood was obtained from full term donors at the Royal London Hospital (UK). The study was approved by the East London Ethical Research committee (REC: 06/Q0604/110) and all participants provided informed consent. Mononuclear cells were isolated by density centrifugation using Ficoll-Paque (GE Healthcare). For in vitro experiments CD34+ cells were enriched using the EasySep CD34+ Selection Kit II (StemCell Technologies). CD34+ cells were cultured in StemSpan SFEM II (StemCell Technologies) supplemented with human SCF (100 ng/µL), human FLT3 ligand (100 ng/µL) and human TPO (100 ng/µL; Peprotech) for 24 h before nucleofection using a 4D-Nucleofector system (Lonza). For in vivo engraftment experiments, we used the EasySep Human Progenitor Cell Enrichment Kit (Stem Cell Technologies) to deplete for lineage marker positive cells and HSPCs were isolated as described in detail previously (48 (link)). After sorting human HSCs (CD34+CD38-) were seeded in StemSpanSFEM (Stem Cell Technologies) supplemented with 100 ng/mL rhFLT-3L, 100 ng/mL rhSCF and 100 ng/mL rhTPO. After 48 h cells were collected for CRISPR editing.
Saunderson E.A., Encabo H.H., Devis J., Rouault-Pierre K., Piganeau M., Bell C.G., Gribben J.G., Bonnet D, & Ficz G. (2023). CRISPR/dCas9 DNA methylation editing is heritable during human hematopoiesis and shapes immune progeny. Proceedings of the National Academy of Sciences of the United States of America, 120(34), e2300224120.
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3

LT-HSC Transduction and Transplantation

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Recombinant retrovirus vectors, MIGR1-IRES-GFP, pMys-Hmga2-IRES-GFP, and pMys-Lin28b-IRES-GFP were provided by A. Iwama (Chiba University, Chuo-ku, Chiba, Japan). LT-HSCs were sorted into 96-well microtiter plates coated with the recombinant fibronectin fragment CH-296 (RetroNectin; Takara Bio Inc.) at 300 cells/well and were incubated in StemSpan (STEMCELL Technologies) supplemented with 100 ng/ml mouse SCF (PeproTech) and 100 ng/ml human Tpo (PeproTech) for 24 h. The cells were transduced with a retrovirus vector at a multiplicity of infection of 800 in the presence of 10 µg/ml protamine sulfate (Sigma-Aldrich) and 1 µg/ml RetroNectin for 24 h. After transduction, cells were further incubated in in StemSpan supplemented with 10 ng/ml SCF and 10 ng/ml TPO. 150 LT-HSCs transduced with the indicated retrovirus were transplanted intravenously into 8-wk-old female C57BL/6 mice irradiated at a dose of 9.5 Gy, together with 2.5 × 105 BM competitor cells from 8-wk-old C57BL/6 mice.
Shimizu T., Kubovcakova L., Nienhold R., Zmajkovic J., Meyer S.C., Hao-Shen H., Geier F., Dirnhofer S., Guglielmelli P., Vannucchi A.M., Feenstra J.D., Kralovics R., Orkin S.H, & Skoda R.C. (2016). Loss of Ezh2 synergizes with JAK2-V617F in initiating myeloproliferative neoplasms and promoting myelofibrosis. The Journal of Experimental Medicine, 213(8), 1479-1496.
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4

Cultured Cell Lines and Patient-Derived AML Cells

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HoxA9-IDH1R132H, HoxA9-IDH1R132C, HoxA9-IDH2R140Q and HoxA9-IDH2172K cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum (FBS), 10 ng/mL of human interleukin 6 (hIL-6), 6 ng/mL of murine interleukin 3 (mIL-3), and 20 ng/mL of murine stem cell factor (mSCF; all from Peprotech) and were incubated at 37°C with 5% CO2 in humidified atmosphere. Patient-derived AML cells, freshly isolated or cryopreserved, were cultured in IMDM medium (Gibco) supplemented with 12,5% FBS (Gibco), 12,5% horse serum (Gibco), 5 µM hydrocortisone, 2,5 mM GlutaMax, 10 ng/ml human FLT3L, 10 ng/ml human TPO, 50 ng/µl human SCF and 10 ng/µl human IL-3 (all from Peprotech) and were incubated at 37°C with 5% CO2 in humidified atmosphere. Treatment was carried out with BAY1436032 or DMSO for indicated time-points and concentrations.
Chaturvedi A., Herbst L., Pusch S., Klett L., Goparaju R., Stichel D., Kaulfuss S., Panknin O., Zimmermann K., Toschi L., Neuhaus R., Haegebarth A., Rehwinkel H., Hess-Stumpp H., Bauser M., Bochtler T., Struys E.A., Sharma A., Bakkali A., Geffers R., Araujo-Cruz M.M., Thol F., Gabdoulline R., Ganser A., Ho A.D., von Deimling A., Rippe K., Heuser M, & Krämer A. (2017). Pan-mutant-IDH1 inhibitor BAY1436032 is highly effective against human IDH1 mutant acute myeloid leukemia in vivo. Leukemia, 31(10), 2020-2028.
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5

Expansion and Xenotransplantation of Human Cord Blood CD34+ Cells

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Human umbilical cord blood-derived CD34+ cells (purchased from Lonza or Stem Cell Technologies) or FACS-purified CD34+CD38-CD90+CD49f+ cells (following staining with APC-CD34 [Biolegend; 560940], PE-CD38 [BD; 347687], FITC-CD90 [Biolegend; 328113], and APC/Cy7-CD49f [Biolegend; 313611]) were cultured in IMDM (Life Technologies) containing 0.1% HSA or 0.1% PVA (Sigma P8136, 363081, or 363146), supplemented with 1% ITSX, 1% P/S/G, 10 mM HEPES. For proliferation assays, 50 cells were seeded per well and supplemented with 10 ng/ml human SCF and 100 ng/ml human TPO (Peprotech). During the cultures, media was refreshed every three days and counted at day-7. For xenograft assays, 2×103 cells were expanded for 7-days before being injected intravenously into sub-lethally-irradiated (1.5 Gy) NOG mice. Human cell chimerism in the PB was calculated at 16-weeks post-transplantation using PE/Cy7-CD45.1 (Biolegend; 110730) and V450-hCD45 antibodies (BD; 560367).
Wilkinson A.C., Ishida R., Kikuchi M., Sudo K., Morita M., Crisostomo R.V., Yamamoto R., Loh K.M., Nakamura Y., Watanabe M., Nakauchi H, & Yamazaki S. (2019). Long-term ex vivo hematopoietic stem cell expansion affords nonconditioned transplantation. Nature, 571(7763), 117-121.
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6

Culturing Leukemia Cell Lines and Patient-Derived AML Cells

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HoxA9-IDH1R132H, HoxA9-IDH1R132C, HoxA9-IDH2R140Q and HoxA9-IDH2172K cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 15% fetal bovine serum (FBS), 10 ng/ml of human interleukin 6 (hIL-6), 6 ng/ml of murine interleukin 3 (mIL-3), and 20 ng/ml of murine stem cell factor (mSCF; all from PeproTech, Hamburg, Germany) and were incubated at 37 °C with 5% CO2 in humidified atmosphere. Patient-derived AML cells, freshly isolated or cryopreserved, were cultured in IMDM medium (Gibco, Karlsruhe, Germany) supplemented with 12.5% FBS (Gibco), 12.5% horse serum (Gibco), 5 μm hydrocortisone, 2.5 mm GlutaMax, 10 ng/ml human FLT3-ligand, 10 ng/ml human TPO, 50 ng/μl human SCF and 10 ng/μl human IL-3 (all from PeproTech) and were incubated at 37 °C with 5% CO2 in humidified atmosphere. Treatment was carried out with BAY1436032 or dimethyl sulfoxide for indicated time points and concentrations.
Chaturvedi A., Herbst L., Pusch S., Klett L., Goparaju R., Stichel D., Kaulfuss S., Panknin O., Zimmermann K., Toschi L., Neuhaus R., Haegebarth A., Rehwinkel H., Hess-Stumpp H., Bauser M., Bochtler T., Struys E.A., Sharma A., Bakkali A., Geffers R., Araujo-Cruz M.M., Thol F., Gabdoulline R., Ganser A., Ho A.D., von Deimling A., Rippe K., Heuser M, & Krämer A. (2017). Pan-mutant-IDH1 inhibitor BAY1436032 is highly effective against human IDH1 mutant acute myeloid leukemia in vivo. Leukemia, 31(10), 2020-2028.
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7

Erythroid Differentiation of CD34+ HSPCs

Cited 2 times
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CD34+ HSPCs were thawed into a maintenance medium consisting of a StemSpan II base (StemCell Technologies), CC100 (StemCell Technologies), 50 ng/mL human TPO (Pepro Tech), and 1% penicillin/streptomycin (Life Technologies).61 (link),62 (link) Cells treated with RNP complexes for enhancer deletions were electroporated 48 hours after thawing and collected 72 hours post-nucleofection. Cells treated with lentivirus were transduced 24 hours after thawing, sorted 72 hours after thawing, and moved to erythroid media 96 hours after thawing.
After the maintenance phase, CD34+ HSPCs were differentiated using the three-phase culture system previously described.63 (link),64 (link) First, a base erythroid medium was created by supplementing IMDM with 2% human AB plasma, 3% human AB serum, 3 U/mL heparin, 10 μg/mL insulin, 200 μg/mL holo-transferrin, and 1% penicillin/streptomycin. From days 1-7 in erythroid media, this base medium was further supplemented with 3 U/mL EPO, 10 ng/mL human SCF, and 1 ng/mL IL-3. From days 7-12, this base medium was further supplemented with 3 U/mL EPO and 10 ng/mL human SCF. After day 12, the base medium was supplemented with 1 mg/mL total of holo-transferrin and 3 U/mL of EPO.
Cato L.D., Li R., Lu H.Y., Yu F., Wissman M., Mkumbe B.S., Ekwattanakit S., Deelen P., Mwita L., Sangeda R., Suksangpleng T., Riolueang S., Bronson P.G., Paul D.S., Kawabata E., Astle W.J., Aguet F., Ardlie K., de Lapuente Portilla A.L., Kang G., Zhang Y., Nouraie S.M., Gordeuk V.R., Gladwin M.T., Garrett M.E., Ashley-Koch A., Telen M.J., Custer B., Kelly S., Dinardo C.L., Sabino E.C., Loureiro P., Carneiro-Proietti A.B., Maximo C., Méndez A., Hammerer-Lercher A., Sheehan V.A., Weiss M.J., Franke L., Nilsson B., Butterworth A.S., Viprakasit V., Nkya S, & Sankaran V.G. (2023). Genetic regulation of fetal hemoglobin across global populations. medRxiv.
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8

Expansion and Clonogenic Assay of CD34+ Cells

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Purified primary CD34+ cells were cultured in Stemspan serum-free medium (Stem Cell Technologies) containing 10 ng/mL human SCF, and 10 ng/mL human TPO (Peprotech) in the presence of metformin. Forty-eight hours after the culture, the cells were transferred to a methylcellulose-based medium (Methocult H4034 Optimum, StemCell Technologies) in triplicate, and colonies were manually counted after 10–14 days.
Yan J.S., Yang M.Y., Zhang X.H., Luo C.H., Du C.K., Jiang Y., Dong X.J., Wang Z.M., Yang L.X., Li Y.D., Xia L, & Lu Y. (2022). Mitochondrial oxidative phosphorylation is dispensable for survival of CD34+ chronic myeloid leukemia stem and progenitor cells. Cell Death & Disease, 13(4), 384.
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9

Megakaryocyte Differentiation from CD34+ Cells

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Frozen human CD34+ cells were obtained from STEMCELL Technologies and differentiated in serum-free expansion medium supplemented with human TPO (100 ng/ml) (PeproTech) (32 (link)). During the culture, megakaryocytic differentiation stages were routinely evaluated by May-Grünwald-Giemsa staining (Sigma) and CD41 levels via flow cytometry. CD34+ cells, MK progenitors, mature MKs, or platelets were collected on day 0, 6, 12, or 15 of the culture, respectively.
Zhang Q., Dong T., Li P, & Wu M.X. (2016). Noninvasive low-level laser therapy for thrombocytopenia. Science translational medicine, 8(349), 349ra101.
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10

Transduction of CD34+ Hematopoietic Stem Cells

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PB CD34+ samples were obtained from healthy donors from plerixafor and G-CSF mobilization. Cells were thawed and plated at 1 × 106 cells/mL on non-tissue culture-treated 96-well plates pre-coated with retronectin (20 μg/mL; Takara Shuzo). Cells were pre-stimulated for 24 h in X-Vivo 15 medium (Lonza) supplemented with 1× glutamine, penicillin, and streptomycin (Gemini Bio-Products), human Flt-3 ligand (50 ng/mL), human SCF (50 ng/mL), human TPO (50 ng/mL), and human IL-3 (20 ng/mL) (cytokines: PeproTech). CD34+ cells were transduced with concentrated viral supernatants at transduction concentrations of 2 × 105 TU/mL, 6 × 105 TU/mL, 2 × 106 TU/mL, and 6 × 106 TU/mL with transduction enhancer Poloxamer 338 (1 mg/mL) (BASF, Ludwigshafen, Germany). At 24 h after transduction, cells were washed and plated under myeloid culture conditions. Cells were cultured for 2 weeks after transduction in basal bone marrow media consisting of Iscove-modified Dulbecco’s medium (Lonza) 1× glutamine, penicillin, and streptomycin (Gemini Bio-Products), 20% fetal bovine serum, and 0.52% BSA (Sigma-Aldrich) supplemented with human SCF (25 ng/mL), human IL-3 (5 ng/mL), and human IL-6 (10 ng/mL).
Hart K.L., Liu B., Brown D., Campo-Fernandez B., Tam K., Orr K., Hollis R.P., Brendel C., Williams D.A, & Kohn D.B. (2024). A novel high-titer, bifunctional lentiviral vector for autologous hematopoietic stem cell gene therapy of sickle cell disease. Molecular Therapy. Methods & Clinical Development, 32(2), 101254.
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