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:

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

Journal: Molecular cell

Article Title: Large DNA Methylation Nadirs Anchor Chromatin Loops Maintaining Hematopoietic Stem Cell Identity

doi: 10.1016/j.molcel.2020.04.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For erythroid progenitor (EP) cells, 0.5-1X10 6 cells magnetically enriched for CD34+ were kept in SFEM II stem cell expansion medium (Stem Cell Technologies) supplemented with 50ng/mL SCF, 25ng/mL Flt3L and 50ng/mL TPO for 5 days.

Techniques: Recombinant, Protease Inhibitor, Transplantation Assay, DNA Methylation Assay, Purification