Journal: PLoS ONE

Article Title: Human intrahepatic ILC2 are IL-13 positive amphiregulin positive and their frequency correlates with model of end stage liver disease score

doi: 10.1371/journal.pone.0188649

Figure Lengend Snippet: (A) ILC gating strategy. Intrahepatic lymphocytes were freshly isolated and gated on the CD45 pos CD3 neg population. The total ILC population was defined as lineage neg and CD127 pos . Of these, ILC1 were defined as CD117 neg CRTH2 neg , ILC2 as CD117 pos/neg CRTH2 pos and ILC3 as CD117 pos CRTH2 neg. (B) Frequency of the total ILC population among the CD3 neg CD45 pos population. (C) Frequencies of the three major ILC subsets in the total intrahepatic ILC in different inflammatory liver diseases. AILD: Autoimmune Liver Diseases (PSC, PBC, AIH); NASH: Nonalcoholic Steatohepatitis; ALD: Alcoholic Liver Disease. (* = p <0.05 by Kruskal Wallis test). (D) Distribution of NKp44 expression among intrahepatic ILC. ILC3 were NKp44 pos (NCR pos ) or NKp44 neg (NCR neg ). (E) Frequencies of NKp44 expression by intrahepatic ILC3 in normal and diseased livers. Summary data are median ± Interquartile range.

Article Snippet: Freshly isolated lymphocytes were stained on ice with antibodies against functional surface markers of interest together with antibodies to identify ILC subsets including: CD3 (UCHT1, BioLegend), CD45 (5B1, Miltenyi Biotec), CD127 (eBioRDR5, eBiosciences) and lineage markers (CD1a (HI149), CD11c (MJ4-27G12), CD14 (TUK4), CD16 (VEP13), CD19 (LT19), CD34 (AC136), CD94 (REA113), CD123 (AC145), BDCA2 (AC144), FceR1 (CRA1) (all from Miltenyi Biotec)) to identify total ILC, CRTH2 (BM16) and CD117 (104D2) (both from BioLegend) to distinguish ILC1 (CRTH2-CD117-), ILC2 (CRTH2+CD117+/-) and ILC3 (CRTH2-CD117+).

Techniques: Isolation, Expressing

Journal: PLoS ONE

Article Title: Human intrahepatic ILC2 are IL-13 positive amphiregulin positive and their frequency correlates with model of end stage liver disease score

doi: 10.1371/journal.pone.0188649

Figure Lengend Snippet: (A) The CD45 pos CD3 neg lineage neg CD127 pos CRTH2 pos ILC2 subset was gated and CD161 and CD69 expressions were analysed. CD161 and CD69 representative overlays and summary data are shown. (B) PGD2 production by human liver. The secretion of PGD2 by normal and inflamed human liver tissue was analysed by ELISA on 24-hour liver tissue supernatants prepared for 1g-tissue/1ml culture medium. Summary data are median ± Interquartile range. (C) Expression of the IL-33 receptor, ST2 was analysed on ILC2. Representative overlay and summary data are shown. In histogram overlays, dotted lines are isotype staining and shaded histograms marker expression.

Article Snippet: Freshly isolated lymphocytes were stained on ice with antibodies against functional surface markers of interest together with antibodies to identify ILC subsets including: CD3 (UCHT1, BioLegend), CD45 (5B1, Miltenyi Biotec), CD127 (eBioRDR5, eBiosciences) and lineage markers (CD1a (HI149), CD11c (MJ4-27G12), CD14 (TUK4), CD16 (VEP13), CD19 (LT19), CD34 (AC136), CD94 (REA113), CD123 (AC145), BDCA2 (AC144), FceR1 (CRA1) (all from Miltenyi Biotec)) to identify total ILC, CRTH2 (BM16) and CD117 (104D2) (both from BioLegend) to distinguish ILC1 (CRTH2-CD117-), ILC2 (CRTH2+CD117+/-) and ILC3 (CRTH2-CD117+).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Staining, Marker

Journal: PLoS ONE

Article Title: Human intrahepatic ILC2 are IL-13 positive amphiregulin positive and their frequency correlates with model of end stage liver disease score

doi: 10.1371/journal.pone.0188649

Figure Lengend Snippet: (A) The CD45 pos CD3 neg lineage neg CD127 pos CRTH2 pos ILC2 subset was gated and expression of the IL-2 receptor α-chain, CD25, was analysed. CD25 representative overlay and summary data in normal and diseased livers is shown. (** = p <0.01 by Mann Whitney test). In the histogram overlay the dotted line represents isotype staining and the shaded histogram reports CD25 staining. (B) Human inflamed liver supernatant was analysed for IL-2, IL-7, IL-9 and IFN-γ. The secretion of cytokines by normal and inflamed human liver tissue was analysed by luminex of 24-hour liver tissue supernatants prepared for 1g of tissue/1ml culture medium. (C) IL-33 production by human liver tissue (Normal and diseased) and (D) IL-33 production by Primary human biliary epithelial (BEC). IL-33 in 24-hour supernatants generated by 1g liver tissue/1ml medium or BEC cells unstimulated, stimulated with IFN-γ and TNF-α or with lipopolysaccharide (LPS) was analysed by ELISA. Summary data are median ± Interquartile range.

Article Snippet: Freshly isolated lymphocytes were stained on ice with antibodies against functional surface markers of interest together with antibodies to identify ILC subsets including: CD3 (UCHT1, BioLegend), CD45 (5B1, Miltenyi Biotec), CD127 (eBioRDR5, eBiosciences) and lineage markers (CD1a (HI149), CD11c (MJ4-27G12), CD14 (TUK4), CD16 (VEP13), CD19 (LT19), CD34 (AC136), CD94 (REA113), CD123 (AC145), BDCA2 (AC144), FceR1 (CRA1) (all from Miltenyi Biotec)) to identify total ILC, CRTH2 (BM16) and CD117 (104D2) (both from BioLegend) to distinguish ILC1 (CRTH2-CD117-), ILC2 (CRTH2+CD117+/-) and ILC3 (CRTH2-CD117+).

Techniques: Expressing, MANN-WHITNEY, Staining, Luminex, Generated, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Human intrahepatic ILC2 are IL-13 positive amphiregulin positive and their frequency correlates with model of end stage liver disease score

doi: 10.1371/journal.pone.0188649

Figure Lengend Snippet: (A) The CD45 pos CD3 neg lineage neg CD127 posi CRTH2 pos ILC2 subset was gated and expressions of CXCR3, CCR6, Very Late Antigen-5 (VLA-5) and Very Late Antigen-6 (VLA-6) were analysed. Representative overlays and summary data in normal and diseased livers are shown (normal livers = open squares; diseased livers = filled squares). Summary data are median ± Interquartile range. In histogram overlays, dotted lines represent isotype staining and shaded histograms show the marker expression. (B) Primary human biliary epithelial cells (BEC), stellate cells and fibroblasts were isolated and stimulated with IFN-γ and TNF-α. Interferon gamma-induced Protein-10 (IP-10) secretion was analysed by ELISA. (* = p<0.05, ** = <0.01 by Paired t- test). Summary data are mean ± SEM. (C) Expressions of CXCR3 and CCR6 by the peripheral blood ILC subsets of normal donors and autoimmune liver disease patients with the condition autoimmune hepatitis (AIH). Summary data are median ± interquartile range. (* = p<0.05 by Mann-Whitney test).

Article Snippet: Freshly isolated lymphocytes were stained on ice with antibodies against functional surface markers of interest together with antibodies to identify ILC subsets including: CD3 (UCHT1, BioLegend), CD45 (5B1, Miltenyi Biotec), CD127 (eBioRDR5, eBiosciences) and lineage markers (CD1a (HI149), CD11c (MJ4-27G12), CD14 (TUK4), CD16 (VEP13), CD19 (LT19), CD34 (AC136), CD94 (REA113), CD123 (AC145), BDCA2 (AC144), FceR1 (CRA1) (all from Miltenyi Biotec)) to identify total ILC, CRTH2 (BM16) and CD117 (104D2) (both from BioLegend) to distinguish ILC1 (CRTH2-CD117-), ILC2 (CRTH2+CD117+/-) and ILC3 (CRTH2-CD117+).

Techniques: Staining, Marker, Expressing, Isolation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Human intrahepatic ILC2 are IL-13 positive amphiregulin positive and their frequency correlates with model of end stage liver disease score

doi: 10.1371/journal.pone.0188649

Figure Lengend Snippet: ( A) Intrahepatic ILC were freshly isolated and stimulated with PMA and Ionomycin for 4 hours and both CD45 pos CD3 neg lineage neg CD127 pos CRTH2 pos ILC2 and CD45 pos CD3 neg lineage neg CD127 pos CRTH2 neg populations were analysed for expressions of IL-4, IL-5, IL-13 and IFN-γ. Representative dot plots and summary data are shown. (B) Immunohistochemistry for amphiregulin in CD3 positive and negative intrahepatic immune cells in human liver. CD3 (Vector Red, red color); Amphiregulin (DAB, Brown color). (C) Amphiregulin expression by intrahepatic ILC2. Amphiregulin representative overlay and summary data in normal and diseased livers are shown. In the histogram overlay the dotted line represents isotype staining and the shaded histogram reports amphiregulin staining.

Article Snippet: Freshly isolated lymphocytes were stained on ice with antibodies against functional surface markers of interest together with antibodies to identify ILC subsets including: CD3 (UCHT1, BioLegend), CD45 (5B1, Miltenyi Biotec), CD127 (eBioRDR5, eBiosciences) and lineage markers (CD1a (HI149), CD11c (MJ4-27G12), CD14 (TUK4), CD16 (VEP13), CD19 (LT19), CD34 (AC136), CD94 (REA113), CD123 (AC145), BDCA2 (AC144), FceR1 (CRA1) (all from Miltenyi Biotec)) to identify total ILC, CRTH2 (BM16) and CD117 (104D2) (both from BioLegend) to distinguish ILC1 (CRTH2-CD117-), ILC2 (CRTH2+CD117+/-) and ILC3 (CRTH2-CD117+).

Techniques: Isolation, Immunohistochemistry, Plasmid Preparation, Expressing, Staining

Journal: Cancer immunology research

Article Title: Combined anti-VEGF and anti–CTLA-4 therapy elicits humoral immunity to galectin-1 which is associated with favorable clinical outcomes

doi: 10.1158/2326-6066.CIR-16-0385

Figure Lengend Snippet: Anti-Gal-1 antibodies isolated from an Ipi-Bev-treated patient abrogate Gal-1 binding to CD45. Anti-Gal-1 antibodies were affinity purified from plasma. Biotinylated HAS-Gal-1 (250 ng/ml) was incubated with a commercial anti-Gal-1 polyclonal antibody (Gal-1 Ab) or control antibody (10 μg/ml), enriched endogenous Gal-1 antibodies (Gal-1 Ig) or normal human IgG (1.98 μg/ml) prior to incubation with coated CD45. The binding of HAS-Gal-1 to CD45 was detected with streptavidin-HRP. Sucrose and lactose were added to the reaction at 5 mM. Data are presented as Mean ± SD of 3 experiments.

Article Snippet: Recombinant human CD45 (R&D Systems) was coated onto 96-well plates (25 ng/well) at 4°C overnight.

Techniques: Isolation, Binding Assay, Affinity Purification, Clinical Proteomics, Incubation, Control

Journal: Biomolecules

Article Title: The Role of Doxycycline and IL-17 in Regenerative Potential of Periodontal Ligament Stem Cells: Implications in Periodontitis

doi: 10.3390/biom13101437

Figure Lengend Snippet: Morphology, immunophenotype and differentiation potential of PDLSCs: ( A ) Adherent PDLSCs with fibroblast-like shape grown in GM under standard conditions for 3 days (scale bars: 50 µM); Florescent images of TRITC-conjugated phalloidin labeled F-actin (red) merged with DAPI (4′,6-diamidino-2-phenylindole) nuclear staining (blue) (scale bars: 10 µM). ( B ) Immunophenotypic characteristics of PDLSCs estimated by flow cytometry. Representative histograms present percentages of cells positive (empty peaks) for mesenchymal markers (CD29, CD44, CD73 and CD105) and hematopoietic markers (CD11b, CD235, CD34 and CD45) in comparison to isotype control (shaded peaks). ( C ) Tri-lineage differentiation potential of PDLSCs. Cells were cultured in growth (GM) and differentiation medium (DM). Osteogenic differentiation was determined by positive staining for ALP activity and extracellular matrix mineralization with Alizarin red; scale bars: 50 m; positive staining of proteoglycans by Safranin O confirmed chondrogenic differentiation of PDLSCs; scale bars: 50 m; Oil Red O staining of intracytoplasmic lipid droplets demonstrated adipogenic differentiation; scale bars: 20 µm.

Article Snippet: Cells were then labeled for 30 min at +4 °C in the dark with monoclonal antibodies specific for human antigens including CD45, CD235a, CD90, CD44H, CD73 (all from R&D Systems, Minneapolis, MN, USA), CD-105, CD29 (Invitrogen, Waltham, MA, USA), CD34 (Dako Cytomation, Glostrup, Denmark) and CD11b (Biosource, Camarillo, CA, USA), each of which was conjugated with either FITC or PE.

Techniques: Labeling, Staining, Flow Cytometry, Comparison, Control, Cell Culture, Activity Assay

Journal: Journal of nanobiotechnology

Article Title: Cerium oxide nanoparticles-carrying human umbilical cord mesenchymal stem cells counteract oxidative damage and facilitate tendon regeneration.

doi: 10.1186/s12951-023-02125-5

Figure Lengend Snippet: Fig. 1 Characterization of hUCMSCs. A Primary hUCMSCs were spindle-shaped under light microscopy. B Flow cytometry analysis showed that 98.5% of cells expressed CD29, 97.4% expressed CD44, 98.6% of cells expressed CD90, and 98.1% of cells expressed CD105. Meanwhile, only 0.48% and 0.28% expressed CD45 and CD34. C Lipid droplets were stained by oil red O after adipogenic differentiation induction of hUCMSCs. Mineralized nodules were stained by Alizarin Red after osteogenic differentiation induction of hUCMSCs. Acid mucopolysaccharide was stained by Alcian Blue after chondrogenic differentiation induction of hUCMSCs

Article Snippet: Briefly, 107 hUCMSCs were subjected to analysis for surface markers CD34 (E-AB-F1143D, Elabscience), CD44 (E-AB-F1100D, Elabscience), CD45 (E-AB-F1137D, Elabscience), CD29 (E-AB-F1049D, Elabscience), CD90 (E-AB-F1167D, Elabscience), and CD105 (E-AB-F1310D, Elabscience) by flow cytometry following the manufacturer’s protocol.

Techniques: Light Microscopy, Flow Cytometry, Staining

Journal: Cell

Article Title: Regenerative Metaplastic Clones in COPD Lung Drive Inflammation and Fibrosis

doi: 10.1016/j.cell.2020.03.047

Figure Lengend Snippet: A. FACS profiles of COPD and control clone libraries using markers established from library scRNAseq and clonal RNAseq including anti-AQP5 (Cluster 1), anti-TRPC6 (Clusters 2+3), and anti-CXCL8 (Cluster 4). B. Histogram compiling FACS quantification data on the relative clone composition of each patient library. C. Histological sections of four-week xenograft of clone libraries from control (SPN-21) showing epithelial cysts devoid of luminal cells. D. Histological sections of four-week xenograft of clone libraries from COPD case (SPN-04) showing epithelial cysts marked by abundant infiltration of CD45+/Ly6G+ leukocytes (insets). E. Histogram depicting the quantification of leukocyte infiltration in xenografts of clone libraries from 11 control and 19 COPD patients based on degree of CD45+ cells in cysts (right). Scale bar, 100 μm. See also Figure S4.

Article Snippet: The sources of primary antibodies used in this study include: Rabbit monoclonal cytokeratin 5 antibody (RM-2106-S; Thermo Fisher, USA); mouse monoclonal human cytokeratin 5 antibody (NCL-L-CK5; Leica Biosystems, Germany); mouse monoclonal CC10 antibody (sc-365992; Santa Cruz Biotechnology, USA); rabbit polyclonal AQP4 antibody (HPA014784; Sigma-Aldrich, USA); rabbit polyclonal SFTPB antibody (HPA034820; Sigma-Aldrich, USA); mouse monoclonal acetylated tubulin T7451 (Sigma-Aldrich, USA); rat monoclonal mouse CD45 antibody 14-0451-85 (Thermo Fisher, USA); mouse monoclonal human CD45 MAB1430-SP (R&D systems, USA); rat monoclonal mouse Ly6G MAB1037 (R&D systems, USA); mouse monoclonal alpha smooth muscle actin antibody (ab7817; Abcam, UK); rabbit polyclonal Involucrin antibody (HPA055211; Sigma-Aldrich, USA); mouse monoclonal Cytokeratin 10 antibody (904301; BioLegend, USA); rabbit polyclonal Muc5B antibody (ab87376; Abcam, UK); rabbit polyclonal Muc5AC antibody (ab78660; Abcam, UK); goat polyclonal E-Cadherin antibody (AF648; R&D Systems, USA).

Techniques: Control

Journal: Cell

Article Title: Regenerative Metaplastic Clones in COPD Lung Drive Inflammation and Fibrosis

doi: 10.1016/j.cell.2020.03.047

Figure Lengend Snippet: A. Histogram depicting most significant (P<1.0e-8) pathways determined by Ingenuity Pathway Analysis of RNAseq differentially expressed genes (FDR<0.05) of patient-matched clones representative of Clusters 1–4. B. Differential expression heatmaps of chemokine, interleukin, and interferon genes among RNAseq DEGs (FDR<0.05) of patient-matched clones representative of Clusters 1–4 (SPN-13). C. H&E on sections through four-week xenografts of patient-matched clones of Clusters 1–4 showing that only Cluster 4 clones are accompanied by abundant intraluminal leukocytes. Scale bar, 100 μm. D. Immunofluorescence micrographs of Cluster 4 xenografts revealing high expression in epithelia of inflammatory mediators including IL33, CXCL8, and IL1B. Scale bar, 50 μm. E. CD45 immunohistochemistry of xenografts derived from Cluster 4 clone grown in vitro to passage 5 and to passage 25. Scale bar, 200 μm. F. Histogram of CXCL8, CCL20, and CXCL10 gene expression in clonal representatives of Clusters 1–4 at in vitro passage 5 and passage 25. See also Figure S5.

Article Snippet: The sources of primary antibodies used in this study include: Rabbit monoclonal cytokeratin 5 antibody (RM-2106-S; Thermo Fisher, USA); mouse monoclonal human cytokeratin 5 antibody (NCL-L-CK5; Leica Biosystems, Germany); mouse monoclonal CC10 antibody (sc-365992; Santa Cruz Biotechnology, USA); rabbit polyclonal AQP4 antibody (HPA014784; Sigma-Aldrich, USA); rabbit polyclonal SFTPB antibody (HPA034820; Sigma-Aldrich, USA); mouse monoclonal acetylated tubulin T7451 (Sigma-Aldrich, USA); rat monoclonal mouse CD45 antibody 14-0451-85 (Thermo Fisher, USA); mouse monoclonal human CD45 MAB1430-SP (R&D systems, USA); rat monoclonal mouse Ly6G MAB1037 (R&D systems, USA); mouse monoclonal alpha smooth muscle actin antibody (ab7817; Abcam, UK); rabbit polyclonal Involucrin antibody (HPA055211; Sigma-Aldrich, USA); mouse monoclonal Cytokeratin 10 antibody (904301; BioLegend, USA); rabbit polyclonal Muc5B antibody (ab87376; Abcam, UK); rabbit polyclonal Muc5AC antibody (ab78660; Abcam, UK); goat polyclonal E-Cadherin antibody (AF648; R&D Systems, USA).

Techniques: Clone Assay, Quantitative Proteomics, Immunofluorescence, Expressing, Immunohistochemistry, Derivative Assay, In Vitro, Gene Expression

Journal: Cell

Article Title: Regenerative Metaplastic Clones in COPD Lung Drive Inflammation and Fibrosis

doi: 10.1016/j.cell.2020.03.047

Figure Lengend Snippet: A. Schematic of clone library generation from pseudoglandular fetal lung and analysis by scRNAseq and xenografting. B. Single cell RNA sequencing of clone library from 13-wk fetal lung yielding tSNE profile (left), integration with three adult control and three COPD libraries (middle), and the fetal subset profile based on the integrated profile (right). C. Histology and indicated marker analysis of xenografts of 13-wk fetal clones corresponding to Clusters 1–4. Cluster 4 xenografts are further assessed by immunohistochemistry with antibodies to CD34 and Ly6G. D. Ratio of qPCR-determined marker expression across Cluster 1–4 clones from COPD, Control, and 13-wk fetal lung. Data represented as mean ± SEM. Cluster 1 markers, AQP5 and NDRG1; Clusters 2 and 3, TRPC6 and ANLN; Cluster 4, CXCL8 and CCL20. E. Percentage composition of Cluster 4 clones across 11 Control and 19 COPD clone libraries. F. Schematic for generating xenografts from defined ratios of Cluster 4 and Cluster 1 cells. G. Histogram of quantification of host inflammatory response to co-xenografts of Cluster 4 and Cluster 1 clones based on CD45 and Ly6G monitoring of cystic infiltration by leukocytes. See also Figure S7 and Table S8.

Article Snippet: The sources of primary antibodies used in this study include: Rabbit monoclonal cytokeratin 5 antibody (RM-2106-S; Thermo Fisher, USA); mouse monoclonal human cytokeratin 5 antibody (NCL-L-CK5; Leica Biosystems, Germany); mouse monoclonal CC10 antibody (sc-365992; Santa Cruz Biotechnology, USA); rabbit polyclonal AQP4 antibody (HPA014784; Sigma-Aldrich, USA); rabbit polyclonal SFTPB antibody (HPA034820; Sigma-Aldrich, USA); mouse monoclonal acetylated tubulin T7451 (Sigma-Aldrich, USA); rat monoclonal mouse CD45 antibody 14-0451-85 (Thermo Fisher, USA); mouse monoclonal human CD45 MAB1430-SP (R&D systems, USA); rat monoclonal mouse Ly6G MAB1037 (R&D systems, USA); mouse monoclonal alpha smooth muscle actin antibody (ab7817; Abcam, UK); rabbit polyclonal Involucrin antibody (HPA055211; Sigma-Aldrich, USA); mouse monoclonal Cytokeratin 10 antibody (904301; BioLegend, USA); rabbit polyclonal Muc5B antibody (ab87376; Abcam, UK); rabbit polyclonal Muc5AC antibody (ab78660; Abcam, UK); goat polyclonal E-Cadherin antibody (AF648; R&D Systems, USA).

Techniques: RNA Sequencing, Control, Marker, Clone Assay, Immunohistochemistry, Expressing

Journal: Cell

Article Title: Regenerative Metaplastic Clones in COPD Lung Drive Inflammation and Fibrosis

doi: 10.1016/j.cell.2020.03.047

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The sources of primary antibodies used in this study include: Rabbit monoclonal cytokeratin 5 antibody (RM-2106-S; Thermo Fisher, USA); mouse monoclonal human cytokeratin 5 antibody (NCL-L-CK5; Leica Biosystems, Germany); mouse monoclonal CC10 antibody (sc-365992; Santa Cruz Biotechnology, USA); rabbit polyclonal AQP4 antibody (HPA014784; Sigma-Aldrich, USA); rabbit polyclonal SFTPB antibody (HPA034820; Sigma-Aldrich, USA); mouse monoclonal acetylated tubulin T7451 (Sigma-Aldrich, USA); rat monoclonal mouse CD45 antibody 14-0451-85 (Thermo Fisher, USA); mouse monoclonal human CD45 MAB1430-SP (R&D systems, USA); rat monoclonal mouse Ly6G MAB1037 (R&D systems, USA); mouse monoclonal alpha smooth muscle actin antibody (ab7817; Abcam, UK); rabbit polyclonal Involucrin antibody (HPA055211; Sigma-Aldrich, USA); mouse monoclonal Cytokeratin 10 antibody (904301; BioLegend, USA); rabbit polyclonal Muc5B antibody (ab87376; Abcam, UK); rabbit polyclonal Muc5AC antibody (ab78660; Abcam, UK); goat polyclonal E-Cadherin antibody (AF648; R&D Systems, USA).

Techniques: Virus, Recombinant, SYBR Green Assay, Membrane, Stem Cell Culture, cDNA Synthesis, Expressing, Microarray, Sequencing, Software

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Contrived samples in HDB. (A) Representative on-chip immunofluorescence image of cells captured with the cartridge of the Hitachi MCA system. Red: CD45; blue: DAPI; green: cytokeratin, white: brightfield image of 8 µm × 100 µm microcavity, imaged at 100×. CTCs are defined as cells that lack CD45 but have a defined DAPI nuclear stain and cytokeratin. (B) Recovery of tumor cell lines spiked into healthy donor blood. The recovery rates of spiked cell lines were 91.5% for H358 and 84.3% for A549. HDB, healthy donor blood; MCA, Micro Cavity Array.

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques: Immunofluorescence, Staining

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Image of CD45 depletion. (A) Shows an MCA cartridge imaged by immunofluorescence where the leukocytes were depleted using CD45 Microbeads prior to enrichment. (B) Shows high levels of co-eluting leukocytes in an MCA cartridge without pre-treatment with CD45 microbeads. MCA, Micro Cavity Array.

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques: Immunofluorescence

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Multivariate analyses

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques:

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Multivariate analyses with CTC count

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques: