easysep human cd34+ selection kit ii  (STEMCELL Technologies Inc)

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Comparison of NSG-Quad and MISTRG-6 humanized mice for modeling circulating and tumor-infiltrating human myeloid cells

doi: 10.1016/j.omtm.2025.101487

Figure Lengend Snippet: NSG-Quad mice support human hematopoietic cell engraftment and multilineage immune cell development (A) Percentage of human CD45 + (hCD45 + ) cells of total CD45 + cells (mouse and human) in the blood at 8 weeks and 10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells. (B) Human immune cell composition in the blood of NSG ( n = 33), NSG-Quad ( n = 34), and MISTRG-6 mice ( n = 32) 8 weeks post-engraftment and in the blood of NSG ( n = 29), NSG-Quad ( n = 25), and MISTRG-6 mice ( n = 16) 10–15 weeks post-engraftment. B cells (CD3 − CD56 − CD33 − CD20 + ), myeloid cells (CD3 − CD56 − CD20 − CD33 + ), NK cells (CD3 − CD56 + ), CD8 + T cells (CD3 + CD4 − CD8 + ), and CD4 + T cells (CD3 + CD8 − CD4 + ). (C) Percentage of human immune cell subsets in the blood 8 weeks post-engraftment (data from B). (D) Percentage of human immune cell subsets in the blood 10–15 weeks post-engraftment (data from B). Data are shown as mean ± SEM. p values were calculated using one-way or two-way ANOVA (A) with Tukey’s multiple comparison test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human CD34 + HSPCs were isolated from cord blood using an EasySep Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies, #17896).

Techniques: Derivative Assay, Comparison

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Comparison of NSG-Quad and MISTRG-6 humanized mice for modeling circulating and tumor-infiltrating human myeloid cells

doi: 10.1016/j.omtm.2025.101487

Figure Lengend Snippet: NSG-Quad mice support the development of human myeloid cells (A) Percentage of human CD45 + (hCD45 + ) cells of total CD45 + cells (mouse and human) in the blood of NSG-Quad +/− mice (heterozygous M-CSF, n = 24) and NSG-Quad +/+ mice (homozygous M-CSF, n = 9) at 10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells. (B) Human immune cell composition in the blood of NSG-Quad +/− mice ( n = 24) and NSG-Quad +/+ mice ( n = 9) 10–15 weeks post-engraftment. (C) Percentage of human immune cell subsets in the blood of NSG-Quad +/− and NSG-Quad +/+ mice 10–15 weeks post-engraftment (data from B). (D) Percentage of human CD45 + (hCD45 + ) cells of total CD45 + cells (mouse and human) in the blood at 8 weeks and 10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells. (E) Human immune cell composition in the blood of NSGS ( n = 11) and NSG-Quad mice ( n = 25) 10–15 weeks post-engraftment. (F) Percentage of human immune cell subsets in the blood of NSGS and NSG-Quad mice 10–15 weeks post-engraftment (data from E). Data are shown as mean ± SEM. p values were calculated using two-tailed, unpaired Student’s t test (A), two-way ANOVA with Tukey’s multiple comparison test (D) and two-tailed, unpaired Mann-Whitney U test (C, F). ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human CD34 + HSPCs were isolated from cord blood using an EasySep Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies, #17896).

Techniques: Derivative Assay, Two Tailed Test, Comparison, MANN-WHITNEY

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Comparison of NSG-Quad and MISTRG-6 humanized mice for modeling circulating and tumor-infiltrating human myeloid cells

doi: 10.1016/j.omtm.2025.101487

Figure Lengend Snippet: NSG-Quad mice promote the development of tumor-infiltrating human macrophages (A) Tumor growth curves in NSG ( n = 9), NSG-Quad ( n = 12), and MISTRG-6 mice ( n = 15) engrafted with SW480 CRC cells and in NSG ( n = 10), NSG-Quad ( n = 4), and MISTRG-6 mice ( n = 14) engrafted with HCT116 CRC cells. (B) Frequency of hCD45 + cells in the tumor of NSG, NSG-Quad, and MISTRG-6 humanized mice engrafted with SW480 or HCT116 CRC cells (10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells; end of experiment). (C) Human immune cell composition in the blood of NSG, NSG-Quad, and MISTRG-6 humanized mice engrafted with SW480 or HCT116 CRC cells (10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells; end of experiment; data from G). (D) Human immune cell composition in the tumor xenografts of NSG ( n = 9, SW480; n = 10, HCT116), NSG-Quad ( n = 10, SW480; n = 3, HCT116), and MISTRG-6 mice ( n = 15, SW480; n = 12, HCT116). (E) Representative IHC pictures show SW480 CRC-infiltrating human CD68 + macrophages in NSG, NSG-Quad, and MISTRG-6 humanized mice. Scale bar: 50 μm. (F) Frequency of human CD68 + macrophages in CRC xenografts of NSG ( n = 11), NSG-Quad ( n = 7), and MISTRG-6 mice ( n = 15) analyzed by IHC. (G) Frequency of CD86 + (M1-like) and CD163 + (M2-like) human CD14 + monocytes in SW480 and HCT116 CRC xenografts of NSG-Quad ( n = 13) and MISTRG-6 mice ( n = 17). (H) Composition of the human CD33 + CD66b − myeloid cell population based on the expression of CD14, CD16, and HLA-DR in SW480 CRC xenografts of NSG-Quad ( n = 8) and MISTRG-6 ( n = 3) mice. Data are shown as mean ± SEM. p values were calculated using one-way or two-way ANOVA (G) with Tukey’s multiple comparison test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Human CD34 + HSPCs were isolated from cord blood using an EasySep Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies, #17896).

Techniques: Derivative Assay, Expressing, Comparison

Journal: Cancer Research Communications

Article Title: PLK1 Inhibition Induces Synthetic Lethality in Fanconi Anemia Pathway–Deficient Acute Myeloid Leukemia

doi: 10.1158/2767-9764.CRC-24-0260

Figure Lengend Snippet: Low-dose volasertib causes selective toxicity in primary AMLs with predicted pathogenic FA pathway mutations. A, Graph represents viability of primary patient CD34 + AML cells treated with 10 nmol/L volasertib for 5 days. P values were calculated by two‐way ANOVA. B, Panel below bar graph indicates predicted pathogenic FA pathway mutations in patient AML samples identified through WES. C, Graph represents Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway functional pathways that were significantly enriched among identified gene mutations ( P value < 0.05). D, Top depicts the position of FANCA residue L1260 within the predicted H12A helix of the C-terminal domain. Protein structure was predicted using AlphaFold Protein Structure Database (version 2022-11-01, RRID: SCR_023662). Bottom shows conservation of L1260 residue (highlighted in yellow) across multiple species. E, Representative Western blot showing FANCA expression in HeLa FANCA- KO cell line transfected with empty vector, FANCA -WT, or FANCA -Mut(L1260P). Representative graph shows the mean ± SEM cell viability (%) of empty ( FANCA- KO), FANCA -WT, and FANCA -Mut(L1260P) HeLa cells treated with MMC for 4 days. P values were calculated by two‐way ANOVA. F, Representative graph shows the mean ± SEM cell viability (%) of empty ( FANCA- KO) and FANCA -WT vs. FANCA -Mut(L1260P) HeLa cells treated with volasertib for 3 days. P values were calculated by two‐way ANOVA. G, Representative Western blot of FAN1 expression in control cells (Scr.Ctrl) vs. FAN1-KD (shFAN1) HeLa cells. Representative graph shows the mean ± SEM cell viability (%) of shFAN1 cells vs. Scr.Ctrl. HeLa cells treated with volasertib for 3 days. For all cell lines, viability was measured by CellTiter-Glo assay. P values were calculated by two‐way ANOVA. All experiments were repeated at least 3 times. ns, not significant.

Article Snippet: CD34 + cells were isolated using EasySep Human Cord Blood CD34 + Selection Kit II (STEMCELL Technologies; 17896) according to the user manual.

Techniques: Functional Assay, Residue, Western Blot, Expressing, Transfection, Plasmid Preparation, Control, Glo Assay