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Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, <t>hematopoietic</t> stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .
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STEMCELL Technologies Inc erythroid maintenance medium stemspan-sfem
Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, <t>hematopoietic</t> stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .
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Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, <t>hematopoietic</t> stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .
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STEMCELL Technologies Inc 150ul stemspan serum-free expansion medium (sfem)
Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, <t>hematopoietic</t> stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .
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STEMCELL Technologies Inc stemspan sfem defined medium
Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, <t>hematopoietic</t> stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .
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Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, hematopoietic stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .

Journal: Frontiers in Immunology

Article Title: Icaritin Induces Anti-tumor Immune Responses in Hepatocellular Carcinoma by Inhibiting Splenic Myeloid-Derived Suppressor Cell Generation

doi: 10.3389/fimmu.2021.609295

Figure Lengend Snippet: Icaritin reduces the accumulation of HSPCs in the spleen of Hepa mice. Spleen weight and total cell number from orthotopic (A) and subcutaneous (B) Hepa mice. Numbers of splenic LSK and LK cells from orthotopic (C) and subcutaneous (D) Hepa mice. Differences between groups were examined for statistical significance by Student's t -test. Data were pooled from two experiments and n = 8 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001. HSPCs, hematopoietic stem and progenitor cells; Hepa, Hepa1-6 cells; ICT, icaritin; Veh, Vehicle; LSK, Lin lo/− Sca-1 + c-Kit hi ; LK, Lin lo/− Sca-1 − c-Kit hi .

Article Snippet: In brief, human CD34 + cells were isolated from the cord blood mononuclear cells using a direct CD34 progenitor cell isolation kit (Miltenyi Biotec), and cultured in hematopoietic stem cell expansion media (StemSpan SFEM; Stem Cell Technologies) supplemented with 100 ng/mL stem cell factor, 100 ng/mL Fms-like tyrosine kinase 3, 100 ng/mL thrombopoietin, and 20 ng/mL IL-3 (R&D Systems) for 9 days ( ).

Techniques:

Icaritin inhibits the generation of human PMN-MDSCs in vitro . Freshly isolated human CD34 + cells from cord blood mononuclear cells were cultured in hematopoietic stem cell expansion media for 8–10 days. The expanded cells were cultured with combined IL-6 and G-CSF in complete medium with 0.1% DMSO (vehicle) or icaritin (2.5 μM) for 3 days. (A) Flow cytometric analysis of CD115 and HLA-DR expression on CD14 + or CD15 + cells. (B) Frequencies of PMN-MDSCs (CD15 + CD115 + ) and M-MDSCs (CD14 + HLA-DR − ). Differences between groups were analyzed using one-way ANOVA, and corrected by Dunnett's test. Data are representative of three experiments. ** P < 0.01. Veh, Vehicle; ICT, Icaritin.

Journal: Frontiers in Immunology

Article Title: Icaritin Induces Anti-tumor Immune Responses in Hepatocellular Carcinoma by Inhibiting Splenic Myeloid-Derived Suppressor Cell Generation

doi: 10.3389/fimmu.2021.609295

Figure Lengend Snippet: Icaritin inhibits the generation of human PMN-MDSCs in vitro . Freshly isolated human CD34 + cells from cord blood mononuclear cells were cultured in hematopoietic stem cell expansion media for 8–10 days. The expanded cells were cultured with combined IL-6 and G-CSF in complete medium with 0.1% DMSO (vehicle) or icaritin (2.5 μM) for 3 days. (A) Flow cytometric analysis of CD115 and HLA-DR expression on CD14 + or CD15 + cells. (B) Frequencies of PMN-MDSCs (CD15 + CD115 + ) and M-MDSCs (CD14 + HLA-DR − ). Differences between groups were analyzed using one-way ANOVA, and corrected by Dunnett's test. Data are representative of three experiments. ** P < 0.01. Veh, Vehicle; ICT, Icaritin.

Article Snippet: In brief, human CD34 + cells were isolated from the cord blood mononuclear cells using a direct CD34 progenitor cell isolation kit (Miltenyi Biotec), and cultured in hematopoietic stem cell expansion media (StemSpan SFEM; Stem Cell Technologies) supplemented with 100 ng/mL stem cell factor, 100 ng/mL Fms-like tyrosine kinase 3, 100 ng/mL thrombopoietin, and 20 ng/mL IL-3 (R&D Systems) for 9 days ( ).

Techniques: In Vitro, Isolation, Cell Culture, Expressing