Journal: PLoS ONE

Article Title: Robust and Highly-Efficient Differentiation of Functional Monocytic Cells from Human Pluripotent Stem Cells under Serum- and Feeder Cell-Free Conditions

doi: 10.1371/journal.pone.0059243

Figure Lengend Snippet: The protocol is composed of 5 steps. CD14-positive cells that are sorted between step-4 are differentiated into dendritic cells by step 5-1 or into macrophages by step 5-2. FL-3: Flt-3 ligand, TPO: Thrombopoietin.

Article Snippet: The CD14 + monocytic lineage-directed cell fraction in supernatant was positively sorted by autoMACS pro (Miltenyi Biotec) with CD14 MicroBeads (Miltenyi Biotec) on days 15–28.

Techniques:

Journal: PLoS ONE

Article Title: Robust and Highly-Efficient Differentiation of Functional Monocytic Cells from Human Pluripotent Stem Cells under Serum- and Feeder Cell-Free Conditions

doi: 10.1371/journal.pone.0059243

Figure Lengend Snippet: (A) Flow cytometric analysis of monocytic lineage cells derived sequentially from pluripotent stem cells. An analysis of adherent cells on day 6 and supernatant cells on day 13 and 18 is shown. (B) May-Giemsa staining of CD14 + monocyte-like cells derived from KhES1 on day 16 (left) and primary human monocytes (right). (C) Esterase staining for CD14 + monocyte-like cells derived from KhES1 on day 16. (D) The percentage of CD14 + cells within the total floating cells derived from KhES1/iPS-201B7 was evaluated from day 13 to day 28. (E) May-Giemsa staining (left) and phase contrast image (right) of mature DCs derived from pluripotent stem cells. (F) Flow cytometric analysis of immature/mature DCs derived from pluripotent stem cells. (G) Phase contrast image and flow cytometric analysis of macrophages derived from pluripotent stem cells.(H) RT-PCR analysis of monocytic lineage cells derived from KhES1/iPS-201B7 clones for expression of monocytic lineage marker genes ( PU.1, c-MAF, TLR4, CCL17 and CCL18 ). Peripheral blood monocytes and peripheral blood monocyte-derived mature DCs were used as positive controls.(A–C, E–G) The data from KhES1-derived cells are shown as representative.

Article Snippet: The CD14 + monocytic lineage-directed cell fraction in supernatant was positively sorted by autoMACS pro (Miltenyi Biotec) with CD14 MicroBeads (Miltenyi Biotec) on days 15–28.

Techniques: Derivative Assay, Staining, Reverse Transcription Polymerase Chain Reaction, Clone Assay, Expressing, Marker

Journal: Frontiers in Microbiology

Article Title: von Willebrand Factor is elevated in HIV patients with a history of thrombosis

doi: 10.3389/fmicb.2015.00180

Figure Lengend Snippet: Plasma levels of LPS ( N = 101), LBP ( N = 130), sCD14 ( N = 130), and vWF antigen ( N = 93) in chronic HIV infected individuals with or without a history of arterial or venous thrombotic disease . Horizontal bar represents median for LPS, LBP, and sCD14; and mean in vWF. Cases had a significantly higher vWF antigen level as compared to controls. Gray, controls without a thrombotic event; red, arterial events; blue, venous events; green, both arterial and venous events. LPS, Lipopolysaccharide; LBP, LPS binding protein; sCD14, soluble CD14; vWF, von Willebrand Factor antigen level. * p = 0.024 for venous events vs. controls.

Article Snippet: LBP was measured using a LBP ELISA kit® (Hycult biotech); soluble CD14 was measured using the Human sCD14 Quantikine ELISA Kit® (R&D systems); vWF antigen levels were measured with an in-house ELISA using DAKOPATTS antibodies and compared to the vWF antigen levels to commercially available pooled plasma from 20 or more otherwise healthy donors (CRYOcheck™).

Techniques: Infection, Binding Assay

Journal: Frontiers in Microbiology

Article Title: von Willebrand Factor is elevated in HIV patients with a history of thrombosis

doi: 10.3389/fmicb.2015.00180

Figure Lengend Snippet: Correlation analysis of sCD14 vs LPS (log-transformed) (A) , sCD14 vs LBP (B) , LBP vs LPS (log-transformed) (C) , vWF vs sCD14 (D) , vWF vs LBP (E) and vWF vs LPS (log-transformed) (F) . A Spearman's Rank-order test was used for analysis. Only sCD14 and LPS (log-transformed) had a statistically significant correlation (A) . There was a weak trend toward positive correlation between vWF and sCD14 (D) . LPS, Lipopolysaccharide; LBP, LPS binding protein; sCD14, soluble CD14; vWF, von Willebrand Factor antigen level.

Article Snippet: LBP was measured using a LBP ELISA kit® (Hycult biotech); soluble CD14 was measured using the Human sCD14 Quantikine ELISA Kit® (R&D systems); vWF antigen levels were measured with an in-house ELISA using DAKOPATTS antibodies and compared to the vWF antigen levels to commercially available pooled plasma from 20 or more otherwise healthy donors (CRYOcheck™).

Techniques: Transformation Assay, Binding Assay