complete blood cell count hemavet 950  (CDC Technologies)

Journal: The Journal of Clinical Investigation

Article Title: Bone marrow–confined IL-6 signaling mediates the progression of myelodysplastic syndromes to acute myeloid leukemia

doi: 10.1172/JCI152673

Figure Lengend Snippet: ( A ) Complete blood cell counts of indicated mice at indicated time points. TWT, Diap1 +/+ Mir146a +/+ Il6 +/+ , n = 11; IL-6 KO, n = 7; DKO, Diap1 –/– Mir146a –/– Il6 +/+ , n = 16; TKO, Diap1 –/– Mir146a –/– Il6 –/– , n = 8. ( B ) Representative spleen images from the indicated mice (left) at 12–14 months of age. The spleen/body weight ratio was further quantified (right). ( C ) Representative histology images of bone marrow and spleen from the indicated mice in B . The reticulin staining reveals increased fibrosis in DKO mice. The arrow and arrowhead indicate mitotic and apoptotic cells, respectively. Scale bars: 100 μm. ( D ) Kaplan-Meier survival analysis of the indicated mice. ( E and F ) In vitro colony-forming unit assay of nucleated cells from the bone marrow (BM), spleen, and peripheral blood (PB) of indicated mice at 12–14 months of age. Representative colonies are shown in E and quantified in F . Scale bars: 500 μm. CFC, colony-forming cell; BMMC, bone marrow mononuclear cell; SPMC, splenic mononuclear cell. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; 1-way ANOVA.

Article Snippet: Complete blood cell counts (Hemavet 950, Drew Scientific) and flow cytometric analysis (BD FACSCanto II) of the peripheral blood were performed at different time points after transplantation to assess chimeras and engraftment.

Techniques: Staining, In Vitro, Colony-forming Unit Assay

Journal: The Journal of Clinical Investigation

Article Title: Bone marrow–confined IL-6 signaling mediates the progression of myelodysplastic syndromes to acute myeloid leukemia

doi: 10.1172/JCI152673

Figure Lengend Snippet: ( A ) Schematic diagram of the transplantation strategies in B – D . ( B ) Kaplan-Meier survival analyses of CD45.1 + mice subjected to transplantation of 2 × 10 6 bone marrow mononuclear cells from the indicated mice. Both the recipient and donor mice were approximately 2 months old at transplantation. ( C and D ) Same as B except 2 × 10 7 splenic mononuclear cells from moribund DKO mice or age-matched wild-type counterparts were used as donor cells. Survival data before ( C ) and after ( D ) 21 days of transplantation are shown. ( E ) Complete blood counts of the recipient mice in D when the mice were 12 weeks post-transplantation. ( F ) Wright-Giemsa staining of peripheral blood smear of mice in E . Arrows indicate blasts. Scale bar: 20 μm. ( G ) Flow cytometric analysis evaluated stem cell surface marker expression in peripheral blood from mice in E . ( H ) Representative flow cytometric profiling of c-Kit + cells in peripheral blood from mice in E . Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; 1-way ANOVA.

Article Snippet: Complete blood cell counts (Hemavet 950, Drew Scientific) and flow cytometric analysis (BD FACSCanto II) of the peripheral blood were performed at different time points after transplantation to assess chimeras and engraftment.

Techniques: Transplantation Assay, Staining, Marker, Expressing

Journal: The Journal of Clinical Investigation

Article Title: Bone marrow–confined IL-6 signaling mediates the progression of myelodysplastic syndromes to acute myeloid leukemia

doi: 10.1172/JCI152673

Figure Lengend Snippet: ( A ) Schematic illustration of bone marrow transplantation. 5 × 10 6 bone marrow cells from 1-year-old DKO mice were transplanted into lethally irradiated 1-year-old CD45.1 + recipient mice. ( B ) Kaplan-Meier survival analysis of the old CD45.1 + recipient mice subjected to transplantation of bone marrow cells from 1-year-old DKO mice as illustrated in A and treated with the indicated reagents once per week. n = 7 in each group. ( C ) Complete blood cell counts of the mice from B given the indicated reagents 1 month after treatment. Data are presented as mean ± SEM. * P < 0.05; 1-way ANOVA. ( D ) Representative H&E staining of the bone marrow, spleen, and liver from the mice in B . Scale bars: 100 μm.

Article Snippet: Complete blood cell counts (Hemavet 950, Drew Scientific) and flow cytometric analysis (BD FACSCanto II) of the peripheral blood were performed at different time points after transplantation to assess chimeras and engraftment.

Techniques: Transplantation Assay, Irradiation, Staining

Journal: The Journal of Clinical Investigation

Article Title: Bone marrow–confined IL-6 signaling mediates the progression of myelodysplastic syndromes to acute myeloid leukemia

doi: 10.1172/JCI152673

Figure Lengend Snippet: ( A ) Cultured MDSL cells were treated with tocilizumab or IgG for 1 hour at indicated concentrations. Cells were then challenged with human recombinant IL-6 (10 ng/mL) for 15 minutes followed by Western blot assay of p-STAT3. Actin was used as a loading control. See complete unedited blots in the supplemental material. ( B ) 1 × 10 4 MDSL cells per well were seeded in a 96-well plate on day 0 in MDSL culture medium with 50 μg/mL tocilizumab or 50 μg/mL human IgG control. Relative cell number was assessed with CCK-8 reagent at indicated time points. ( C ) 1 × 10 6 MDSL cells were transplanted into sublethally irradiated NSG recipient mice. Ten days after transplantation, mice were subjected to weekly tocilizumab (TCZ) or human IgG (8 mg/kg) by i.p. administration. Engraftment was evaluated 60 days after transplantation via flow cytometry assays of hCD45 + mononuclear cells in the peripheral blood. n = 5 in each group. ( D ) Quantification of the percentage of hCD45 + cells in C . ( E and F ) Colony-forming unit (CFU) assays in normal ( E ) and high-risk MDS patient ( F ) bone marrow–derived CD34 + cells. 1 × 10 3 normal ( E ) or 2 × 10 3 patient CD34 + cells ( F ) were seeded in MethoCult medium supplemented with human IgG or tocilizumab (50 μg/mL) on day 0. The number of colonies was assessed on day 14. Triplicate assay colonies were independently identified by 2 individuals. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001; 2-tailed Student’s t test. E, G, M, GM, and GEMM represent BFU/CFU-E, CFU-G, CFU-M, CFU-GM, and CFU-GEMM, respectively, in both E and F .

Article Snippet: Complete blood cell counts (Hemavet 950, Drew Scientific) and flow cytometric analysis (BD FACSCanto II) of the peripheral blood were performed at different time points after transplantation to assess chimeras and engraftment.

Techniques: Cell Culture, Recombinant, Western Blot, Control, CCK-8 Assay, Irradiation, Transplantation Assay, Flow Cytometry, Derivative Assay