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granulocyte isolation beads miltenyi biotec  (Miltenyi Biotec)


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    Miltenyi Biotec granulocyte isolation beads miltenyi biotec
    Granulocyte Isolation Beads Miltenyi Biotec, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd56+microbeads/CD56+MicroBeads%2C+human+-+lyophilized/10__1016_slash_j__isci__2026__116736-639-184-187
    Average 96 stars, based on 24 article reviews
    granulocyte isolation beads miltenyi biotec - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Flow Cytometry:

    Article Title: Hypoxia restricts the generation of BCG-responsive polycytotoxic T cells.
    Article Snippet: .. The following fluorochrome-conjugated antibodies were used for flow cytometry: CD3-PerCP (clone SK7, BD Biosciences), CD8α–APC (clone RPA-T8, BD Pharmingen), CD8β (clone 2ST8.5H7, BD Biosciences), CD4–BV421 (clone RPA-T4, BioLegend), IFN-γ-PE (clone 45–15, Miltenyi), granulysin-Alexa Fluor 488 (clone RB1, BD Biosciences), perforin-PE-Cy7 (clone B–D48, BioLegend), granzyme B-Pacific Blue (clone GB11, BioLegend), and viability dye (Amcyan, Thermo Fisher Scientific).The following reagents were used: Ficoll-Paque PLUS (Cytiva), Macrophage Serum-Free Medium (Gibco), recombinant human GM-CSF and IL-4 (both from R&D), IFN-γ (R&D), LPS (Sigma-Aldrich), CD14 and CD56 MicroBeads (Miltenyi Biotec), IL-2 T6 supernatant (gift of Dr. F. Sallusto, Bellinzona, Switzerland), paraformaldehyde (SigmaAldrich), Perm/Wash buffer (BD Biosciences), and brefeldin A (SigmaAldrich). .. Cytometric bead array kits for CD8/NK profiling (13-plex, V02) were obtained from BioLegend.

    Recombinase Polymerase Amplification:

    Article Title: Hypoxia restricts the generation of BCG-responsive polycytotoxic T cells.
    Article Snippet: .. The following fluorochrome-conjugated antibodies were used for flow cytometry: CD3-PerCP (clone SK7, BD Biosciences), CD8α–APC (clone RPA-T8, BD Pharmingen), CD8β (clone 2ST8.5H7, BD Biosciences), CD4–BV421 (clone RPA-T4, BioLegend), IFN-γ-PE (clone 45–15, Miltenyi), granulysin-Alexa Fluor 488 (clone RB1, BD Biosciences), perforin-PE-Cy7 (clone B–D48, BioLegend), granzyme B-Pacific Blue (clone GB11, BioLegend), and viability dye (Amcyan, Thermo Fisher Scientific).The following reagents were used: Ficoll-Paque PLUS (Cytiva), Macrophage Serum-Free Medium (Gibco), recombinant human GM-CSF and IL-4 (both from R&D), IFN-γ (R&D), LPS (Sigma-Aldrich), CD14 and CD56 MicroBeads (Miltenyi Biotec), IL-2 T6 supernatant (gift of Dr. F. Sallusto, Bellinzona, Switzerland), paraformaldehyde (SigmaAldrich), Perm/Wash buffer (BD Biosciences), and brefeldin A (SigmaAldrich). .. Cytometric bead array kits for CD8/NK profiling (13-plex, V02) were obtained from BioLegend.

    Recombinant:

    Article Title: Hypoxia restricts the generation of BCG-responsive polycytotoxic T cells.
    Article Snippet: .. The following fluorochrome-conjugated antibodies were used for flow cytometry: CD3-PerCP (clone SK7, BD Biosciences), CD8α–APC (clone RPA-T8, BD Pharmingen), CD8β (clone 2ST8.5H7, BD Biosciences), CD4–BV421 (clone RPA-T4, BioLegend), IFN-γ-PE (clone 45–15, Miltenyi), granulysin-Alexa Fluor 488 (clone RB1, BD Biosciences), perforin-PE-Cy7 (clone B–D48, BioLegend), granzyme B-Pacific Blue (clone GB11, BioLegend), and viability dye (Amcyan, Thermo Fisher Scientific).The following reagents were used: Ficoll-Paque PLUS (Cytiva), Macrophage Serum-Free Medium (Gibco), recombinant human GM-CSF and IL-4 (both from R&D), IFN-γ (R&D), LPS (Sigma-Aldrich), CD14 and CD56 MicroBeads (Miltenyi Biotec), IL-2 T6 supernatant (gift of Dr. F. Sallusto, Bellinzona, Switzerland), paraformaldehyde (SigmaAldrich), Perm/Wash buffer (BD Biosciences), and brefeldin A (SigmaAldrich). .. Cytometric bead array kits for CD8/NK profiling (13-plex, V02) were obtained from BioLegend.

    Isolation:

    Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
    Article Snippet: .. Human NK cells were isolated by CD56 Microbeads, human (Miltenyi Biotec), followed by activating and culturing in complete RPMI 1640 medium containing hIL-2 (100 U/mL) and human IL-15 (20 ng/mL) for 24 h. Preparation of Ldha-WT and Ldha-KO NK cells NK cells were isolated from splenocytes of CRISPR–Cas9 knockin mice. .. To generate Ldha-WT NK and Ldha-KO NK cells, NK cells were isolated in the same method described above and activated with IL-2 (50 ng/mL) contained complete RPMI-1640 medium for 24 h, followed by transducing with retrovirus containing scramble control guide RNA or Ldha-targeting gRNA on 6-well plate coated with protamine (10 μg/mL, Thermo Fisher Scientific).

    Article Title: Strength in Unity: a Dual Strategy to Restore NK Cell Cytotoxicity against Pancreatic Ductal Adenocarcinoma
    Article Snippet: F(spontaneous) is the fluorescence released from target cells in the absence of effector cells, and F(maximum) represents the fluorescence released after total cell lysis induced by addition of 1% Triton X100 (Sigma-Aldrich, #X100-1L). .. Purified NK cells (300,000 cells/condition) isolated from healthy donors PBMCs (Red Cross Luxembourg) using positive selection with CD56 Microbeads (Miltenyi Biotec,# 130-097-042) or PDAC patient PBMCs (BioIVT, #HUMANPBMCCL-0120962) (400,000 cells/condition) were incubated with NaMiX, rhu-IL15 or control medium for 48 hours similarly to the activation assays described above. .. After pre-incubation, BxPC-3 cells were stained with CellTrace Violet Cell Proliferation kit (Thermo Fisher Scientific, #C34571) following the manufacturer’s instructions and co-incubated with NK cells for 5 hours at an E:T ratio 1:1 or 2:1 for purified NK cells and 10:1 for PDAC PBMCs.

    Article Title: Feeder-free ex vivo expansion of cord blood-derived natural killer cells for enhanced proliferation and functional maturation
    Article Snippet: Prior to CD56 + cell isolation by magnetic-activated cell sorting (MACS), a pre-enrichment characterization was performed to determine the percentage of CD3 − CD56 + cells by flow cytometry (CD3 PerCP-Cy5.5 (BioLegend) and CD56 PE (BioLegend)). .. CD56 + cells were isolated using MACS with CD56 MicroBeads (Miltenyi Biotec), following the manufacturer instructions, and MACS efficiency was calculated using Eq. .. Briefly, MNC(CB) were resuspended in PBS supplemented with 0.5% (v/v) bovine serum albumin (Sigma-Aldrich) and 2 mM EDTA (MACS Buffer).

    Article Title: Feeder-free ex vivo expansion of cord blood-derived natural killer cells for enhanced proliferation and functional maturation.
    Article Snippet: Prior to CD56+ cell isolation by magnetic-activated cell sorting (MACS), a preenrichment characterization was performed to determine the percentage of CD3CD56+ cells by flow cytometry (CD3 PerCP-Cy5.5 (BioLegend) and CD56 PE (BioLegend)). .. CD56+ cells were isolated using MACS with CD56 MicroBeads (Miltenyi Biotec), following the manufacturer instructions, and MACS efficiency was calculated using Equation 1. .. Briefly, MNC(CB) were resuspended in PBS supplemented with 0.5% (v/v) bovine serum albumin (Sigma-Aldrich) and 2 mM EDTA (MACS Buffer).

    CRISPR:

    Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
    Article Snippet: .. Human NK cells were isolated by CD56 Microbeads, human (Miltenyi Biotec), followed by activating and culturing in complete RPMI 1640 medium containing hIL-2 (100 U/mL) and human IL-15 (20 ng/mL) for 24 h. Preparation of Ldha-WT and Ldha-KO NK cells NK cells were isolated from splenocytes of CRISPR–Cas9 knockin mice. .. To generate Ldha-WT NK and Ldha-KO NK cells, NK cells were isolated in the same method described above and activated with IL-2 (50 ng/mL) contained complete RPMI-1640 medium for 24 h, followed by transducing with retrovirus containing scramble control guide RNA or Ldha-targeting gRNA on 6-well plate coated with protamine (10 μg/mL, Thermo Fisher Scientific).

    Knock-In:

    Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
    Article Snippet: .. Human NK cells were isolated by CD56 Microbeads, human (Miltenyi Biotec), followed by activating and culturing in complete RPMI 1640 medium containing hIL-2 (100 U/mL) and human IL-15 (20 ng/mL) for 24 h. Preparation of Ldha-WT and Ldha-KO NK cells NK cells were isolated from splenocytes of CRISPR–Cas9 knockin mice. .. To generate Ldha-WT NK and Ldha-KO NK cells, NK cells were isolated in the same method described above and activated with IL-2 (50 ng/mL) contained complete RPMI-1640 medium for 24 h, followed by transducing with retrovirus containing scramble control guide RNA or Ldha-targeting gRNA on 6-well plate coated with protamine (10 μg/mL, Thermo Fisher Scientific).

    Purification:

    Article Title: Strength in Unity: a Dual Strategy to Restore NK Cell Cytotoxicity against Pancreatic Ductal Adenocarcinoma
    Article Snippet: F(spontaneous) is the fluorescence released from target cells in the absence of effector cells, and F(maximum) represents the fluorescence released after total cell lysis induced by addition of 1% Triton X100 (Sigma-Aldrich, #X100-1L). .. Purified NK cells (300,000 cells/condition) isolated from healthy donors PBMCs (Red Cross Luxembourg) using positive selection with CD56 Microbeads (Miltenyi Biotec,# 130-097-042) or PDAC patient PBMCs (BioIVT, #HUMANPBMCCL-0120962) (400,000 cells/condition) were incubated with NaMiX, rhu-IL15 or control medium for 48 hours similarly to the activation assays described above. .. After pre-incubation, BxPC-3 cells were stained with CellTrace Violet Cell Proliferation kit (Thermo Fisher Scientific, #C34571) following the manufacturer’s instructions and co-incubated with NK cells for 5 hours at an E:T ratio 1:1 or 2:1 for purified NK cells and 10:1 for PDAC PBMCs.

    Article Title: Hypoxia restricts the generation of BCG-responsive polycytotoxic T cells.
    Article Snippet: .. To isolate autologous T lymphocytes, untouched cells from the monocyte purification were collected, and natural killer (NK) cells were depleted using CD56 Microbeads (Miltenyi Biotec). ..

    Selection:

    Article Title: Strength in Unity: a Dual Strategy to Restore NK Cell Cytotoxicity against Pancreatic Ductal Adenocarcinoma
    Article Snippet: F(spontaneous) is the fluorescence released from target cells in the absence of effector cells, and F(maximum) represents the fluorescence released after total cell lysis induced by addition of 1% Triton X100 (Sigma-Aldrich, #X100-1L). .. Purified NK cells (300,000 cells/condition) isolated from healthy donors PBMCs (Red Cross Luxembourg) using positive selection with CD56 Microbeads (Miltenyi Biotec,# 130-097-042) or PDAC patient PBMCs (BioIVT, #HUMANPBMCCL-0120962) (400,000 cells/condition) were incubated with NaMiX, rhu-IL15 or control medium for 48 hours similarly to the activation assays described above. .. After pre-incubation, BxPC-3 cells were stained with CellTrace Violet Cell Proliferation kit (Thermo Fisher Scientific, #C34571) following the manufacturer’s instructions and co-incubated with NK cells for 5 hours at an E:T ratio 1:1 or 2:1 for purified NK cells and 10:1 for PDAC PBMCs.

    Incubation:

    Article Title: Strength in Unity: a Dual Strategy to Restore NK Cell Cytotoxicity against Pancreatic Ductal Adenocarcinoma
    Article Snippet: F(spontaneous) is the fluorescence released from target cells in the absence of effector cells, and F(maximum) represents the fluorescence released after total cell lysis induced by addition of 1% Triton X100 (Sigma-Aldrich, #X100-1L). .. Purified NK cells (300,000 cells/condition) isolated from healthy donors PBMCs (Red Cross Luxembourg) using positive selection with CD56 Microbeads (Miltenyi Biotec,# 130-097-042) or PDAC patient PBMCs (BioIVT, #HUMANPBMCCL-0120962) (400,000 cells/condition) were incubated with NaMiX, rhu-IL15 or control medium for 48 hours similarly to the activation assays described above. .. After pre-incubation, BxPC-3 cells were stained with CellTrace Violet Cell Proliferation kit (Thermo Fisher Scientific, #C34571) following the manufacturer’s instructions and co-incubated with NK cells for 5 hours at an E:T ratio 1:1 or 2:1 for purified NK cells and 10:1 for PDAC PBMCs.

    Control:

    Article Title: Strength in Unity: a Dual Strategy to Restore NK Cell Cytotoxicity against Pancreatic Ductal Adenocarcinoma
    Article Snippet: F(spontaneous) is the fluorescence released from target cells in the absence of effector cells, and F(maximum) represents the fluorescence released after total cell lysis induced by addition of 1% Triton X100 (Sigma-Aldrich, #X100-1L). .. Purified NK cells (300,000 cells/condition) isolated from healthy donors PBMCs (Red Cross Luxembourg) using positive selection with CD56 Microbeads (Miltenyi Biotec,# 130-097-042) or PDAC patient PBMCs (BioIVT, #HUMANPBMCCL-0120962) (400,000 cells/condition) were incubated with NaMiX, rhu-IL15 or control medium for 48 hours similarly to the activation assays described above. .. After pre-incubation, BxPC-3 cells were stained with CellTrace Violet Cell Proliferation kit (Thermo Fisher Scientific, #C34571) following the manufacturer’s instructions and co-incubated with NK cells for 5 hours at an E:T ratio 1:1 or 2:1 for purified NK cells and 10:1 for PDAC PBMCs.

    Activation Assay:

    Article Title: Strength in Unity: a Dual Strategy to Restore NK Cell Cytotoxicity against Pancreatic Ductal Adenocarcinoma
    Article Snippet: F(spontaneous) is the fluorescence released from target cells in the absence of effector cells, and F(maximum) represents the fluorescence released after total cell lysis induced by addition of 1% Triton X100 (Sigma-Aldrich, #X100-1L). .. Purified NK cells (300,000 cells/condition) isolated from healthy donors PBMCs (Red Cross Luxembourg) using positive selection with CD56 Microbeads (Miltenyi Biotec,# 130-097-042) or PDAC patient PBMCs (BioIVT, #HUMANPBMCCL-0120962) (400,000 cells/condition) were incubated with NaMiX, rhu-IL15 or control medium for 48 hours similarly to the activation assays described above. .. After pre-incubation, BxPC-3 cells were stained with CellTrace Violet Cell Proliferation kit (Thermo Fisher Scientific, #C34571) following the manufacturer’s instructions and co-incubated with NK cells for 5 hours at an E:T ratio 1:1 or 2:1 for purified NK cells and 10:1 for PDAC PBMCs.

    Magnetic Cell Separation:

    Article Title: Feeder-free ex vivo expansion of cord blood-derived natural killer cells for enhanced proliferation and functional maturation
    Article Snippet: Prior to CD56 + cell isolation by magnetic-activated cell sorting (MACS), a pre-enrichment characterization was performed to determine the percentage of CD3 − CD56 + cells by flow cytometry (CD3 PerCP-Cy5.5 (BioLegend) and CD56 PE (BioLegend)). .. CD56 + cells were isolated using MACS with CD56 MicroBeads (Miltenyi Biotec), following the manufacturer instructions, and MACS efficiency was calculated using Eq. .. Briefly, MNC(CB) were resuspended in PBS supplemented with 0.5% (v/v) bovine serum albumin (Sigma-Aldrich) and 2 mM EDTA (MACS Buffer).

    Article Title: Feeder-free ex vivo expansion of cord blood-derived natural killer cells for enhanced proliferation and functional maturation.
    Article Snippet: Prior to CD56+ cell isolation by magnetic-activated cell sorting (MACS), a preenrichment characterization was performed to determine the percentage of CD3CD56+ cells by flow cytometry (CD3 PerCP-Cy5.5 (BioLegend) and CD56 PE (BioLegend)). .. CD56+ cells were isolated using MACS with CD56 MicroBeads (Miltenyi Biotec), following the manufacturer instructions, and MACS efficiency was calculated using Equation 1. .. Briefly, MNC(CB) were resuspended in PBS supplemented with 0.5% (v/v) bovine serum albumin (Sigma-Aldrich) and 2 mM EDTA (MACS Buffer).



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    Characterization of <t>NCAM1</t> + EpCAM + HPCs in the expanded biliary clusters in BA. (a) Enrichment score of HPCs gene set in each biliary cluster. Comparisons among multiple clusters were performed using the Kruskal‐Wallis test, with a significance level of p < 0.001. (b) Venn diagram of DEGs showing higher expression in cluster 17 than other three biliary clusters (avg_log 2 FC>1), the DEGs showing higher expression in BA than the control group across all four biliary clusters (avg_log 2 FC>1), and the membrane protein database. (c) Gene density plot of NCAM1 in biliary clusters. (d): The proportion of NCAM1 positive cells of BA, Ctrl and external single‐cell dataset. P value was calculated by Chi‐squared test. (e): The proportion of NCAM1 + cells in EpCAM + cells of BA, Ctrl and normal cholangiocyte dataset ( GSE185477 contains four healthy human liver caudate lobes). P value was calculated by Chi‐squared test. (f): The expression of NCAM1 in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (g): The expression of NCAM1 in BA (n = 38) and Ctrl liver tissues ( n = 21). P value was calculated by unpaired t test. (h) Immunohistochemical staining of NCAM1 on paraffin sections of livers from Ctrl, BA with mild liver fibrosis (BA‐FM) and BA with severe liver fibrosis (BA‐FS). Scale bar: 100 µm. (i) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA ( n = 42) and Ctrl (n = 10). P value was calculated by unpaired t test. (j) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA‐FM ( n = 30) and BA‐FS ( n = 12). P value was calculated by unpaired t test. (k) Morphology of NCAM1 + EpCAM + cells derived organoids and NCAM1 − EpCAM + cells derived organoids in BA (The left side of each group, scale bar: 200 µm). Regions highlighted with dotted boxes were magnified and shown on the right (scale bar: 50 µm). Calcein AM staining (Green) of the organoids were shown. (l) Volcano plot of DEGs of organoids derived from NCAM1 + EpCAM + cells ( n = 3) and NCAM1 − EpCAM + cells ( n = 4). (m) Bar chart showing transcriptional activity of TFs implicated in biliary and hepatic lineage. Red color indicated increased activity, while blue color indicated decreased activity in NCAM1 + EpCAM + cells derived organoids.
    Ls Columns, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of <t>NCAM1</t> + EpCAM + HPCs in the expanded biliary clusters in BA. (a) Enrichment score of HPCs gene set in each biliary cluster. Comparisons among multiple clusters were performed using the Kruskal‐Wallis test, with a significance level of p < 0.001. (b) Venn diagram of DEGs showing higher expression in cluster 17 than other three biliary clusters (avg_log 2 FC>1), the DEGs showing higher expression in BA than the control group across all four biliary clusters (avg_log 2 FC>1), and the membrane protein database. (c) Gene density plot of NCAM1 in biliary clusters. (d): The proportion of NCAM1 positive cells of BA, Ctrl and external single‐cell dataset. P value was calculated by Chi‐squared test. (e): The proportion of NCAM1 + cells in EpCAM + cells of BA, Ctrl and normal cholangiocyte dataset ( GSE185477 contains four healthy human liver caudate lobes). P value was calculated by Chi‐squared test. (f): The expression of NCAM1 in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (g): The expression of NCAM1 in BA (n = 38) and Ctrl liver tissues ( n = 21). P value was calculated by unpaired t test. (h) Immunohistochemical staining of NCAM1 on paraffin sections of livers from Ctrl, BA with mild liver fibrosis (BA‐FM) and BA with severe liver fibrosis (BA‐FS). Scale bar: 100 µm. (i) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA ( n = 42) and Ctrl (n = 10). P value was calculated by unpaired t test. (j) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA‐FM ( n = 30) and BA‐FS ( n = 12). P value was calculated by unpaired t test. (k) Morphology of NCAM1 + EpCAM + cells derived organoids and NCAM1 − EpCAM + cells derived organoids in BA (The left side of each group, scale bar: 200 µm). Regions highlighted with dotted boxes were magnified and shown on the right (scale bar: 50 µm). Calcein AM staining (Green) of the organoids were shown. (l) Volcano plot of DEGs of organoids derived from NCAM1 + EpCAM + cells ( n = 3) and NCAM1 − EpCAM + cells ( n = 4). (m) Bar chart showing transcriptional activity of TFs implicated in biliary and hepatic lineage. Red color indicated increased activity, while blue color indicated decreased activity in NCAM1 + EpCAM + cells derived organoids.
    Human, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of <t>NCAM1</t> + EpCAM + HPCs in the expanded biliary clusters in BA. (a) Enrichment score of HPCs gene set in each biliary cluster. Comparisons among multiple clusters were performed using the Kruskal‐Wallis test, with a significance level of p < 0.001. (b) Venn diagram of DEGs showing higher expression in cluster 17 than other three biliary clusters (avg_log 2 FC>1), the DEGs showing higher expression in BA than the control group across all four biliary clusters (avg_log 2 FC>1), and the membrane protein database. (c) Gene density plot of NCAM1 in biliary clusters. (d): The proportion of NCAM1 positive cells of BA, Ctrl and external single‐cell dataset. P value was calculated by Chi‐squared test. (e): The proportion of NCAM1 + cells in EpCAM + cells of BA, Ctrl and normal cholangiocyte dataset ( GSE185477 contains four healthy human liver caudate lobes). P value was calculated by Chi‐squared test. (f): The expression of NCAM1 in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (g): The expression of NCAM1 in BA (n = 38) and Ctrl liver tissues ( n = 21). P value was calculated by unpaired t test. (h) Immunohistochemical staining of NCAM1 on paraffin sections of livers from Ctrl, BA with mild liver fibrosis (BA‐FM) and BA with severe liver fibrosis (BA‐FS). Scale bar: 100 µm. (i) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA ( n = 42) and Ctrl (n = 10). P value was calculated by unpaired t test. (j) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA‐FM ( n = 30) and BA‐FS ( n = 12). P value was calculated by unpaired t test. (k) Morphology of NCAM1 + EpCAM + cells derived organoids and NCAM1 − EpCAM + cells derived organoids in BA (The left side of each group, scale bar: 200 µm). Regions highlighted with dotted boxes were magnified and shown on the right (scale bar: 50 µm). Calcein AM staining (Green) of the organoids were shown. (l) Volcano plot of DEGs of organoids derived from NCAM1 + EpCAM + cells ( n = 3) and NCAM1 − EpCAM + cells ( n = 4). (m) Bar chart showing transcriptional activity of TFs implicated in biliary and hepatic lineage. Red color indicated increased activity, while blue color indicated decreased activity in NCAM1 + EpCAM + cells derived organoids.
    Macs Cd56 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of <t>NCAM1</t> + EpCAM + HPCs in the expanded biliary clusters in BA. (a) Enrichment score of HPCs gene set in each biliary cluster. Comparisons among multiple clusters were performed using the Kruskal‐Wallis test, with a significance level of p < 0.001. (b) Venn diagram of DEGs showing higher expression in cluster 17 than other three biliary clusters (avg_log 2 FC>1), the DEGs showing higher expression in BA than the control group across all four biliary clusters (avg_log 2 FC>1), and the membrane protein database. (c) Gene density plot of NCAM1 in biliary clusters. (d): The proportion of NCAM1 positive cells of BA, Ctrl and external single‐cell dataset. P value was calculated by Chi‐squared test. (e): The proportion of NCAM1 + cells in EpCAM + cells of BA, Ctrl and normal cholangiocyte dataset ( GSE185477 contains four healthy human liver caudate lobes). P value was calculated by Chi‐squared test. (f): The expression of NCAM1 in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (g): The expression of NCAM1 in BA (n = 38) and Ctrl liver tissues ( n = 21). P value was calculated by unpaired t test. (h) Immunohistochemical staining of NCAM1 on paraffin sections of livers from Ctrl, BA with mild liver fibrosis (BA‐FM) and BA with severe liver fibrosis (BA‐FS). Scale bar: 100 µm. (i) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA ( n = 42) and Ctrl (n = 10). P value was calculated by unpaired t test. (j) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA‐FM ( n = 30) and BA‐FS ( n = 12). P value was calculated by unpaired t test. (k) Morphology of NCAM1 + EpCAM + cells derived organoids and NCAM1 − EpCAM + cells derived organoids in BA (The left side of each group, scale bar: 200 µm). Regions highlighted with dotted boxes were magnified and shown on the right (scale bar: 50 µm). Calcein AM staining (Green) of the organoids were shown. (l) Volcano plot of DEGs of organoids derived from NCAM1 + EpCAM + cells ( n = 3) and NCAM1 − EpCAM + cells ( n = 4). (m) Bar chart showing transcriptional activity of TFs implicated in biliary and hepatic lineage. Red color indicated increased activity, while blue color indicated decreased activity in NCAM1 + EpCAM + cells derived organoids.
    Cd56 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of NCAM1 + EpCAM + HPCs in the expanded biliary clusters in BA. (a) Enrichment score of HPCs gene set in each biliary cluster. Comparisons among multiple clusters were performed using the Kruskal‐Wallis test, with a significance level of p < 0.001. (b) Venn diagram of DEGs showing higher expression in cluster 17 than other three biliary clusters (avg_log 2 FC>1), the DEGs showing higher expression in BA than the control group across all four biliary clusters (avg_log 2 FC>1), and the membrane protein database. (c) Gene density plot of NCAM1 in biliary clusters. (d): The proportion of NCAM1 positive cells of BA, Ctrl and external single‐cell dataset. P value was calculated by Chi‐squared test. (e): The proportion of NCAM1 + cells in EpCAM + cells of BA, Ctrl and normal cholangiocyte dataset ( GSE185477 contains four healthy human liver caudate lobes). P value was calculated by Chi‐squared test. (f): The expression of NCAM1 in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (g): The expression of NCAM1 in BA (n = 38) and Ctrl liver tissues ( n = 21). P value was calculated by unpaired t test. (h) Immunohistochemical staining of NCAM1 on paraffin sections of livers from Ctrl, BA with mild liver fibrosis (BA‐FM) and BA with severe liver fibrosis (BA‐FS). Scale bar: 100 µm. (i) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA ( n = 42) and Ctrl (n = 10). P value was calculated by unpaired t test. (j) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA‐FM ( n = 30) and BA‐FS ( n = 12). P value was calculated by unpaired t test. (k) Morphology of NCAM1 + EpCAM + cells derived organoids and NCAM1 − EpCAM + cells derived organoids in BA (The left side of each group, scale bar: 200 µm). Regions highlighted with dotted boxes were magnified and shown on the right (scale bar: 50 µm). Calcein AM staining (Green) of the organoids were shown. (l) Volcano plot of DEGs of organoids derived from NCAM1 + EpCAM + cells ( n = 3) and NCAM1 − EpCAM + cells ( n = 4). (m) Bar chart showing transcriptional activity of TFs implicated in biliary and hepatic lineage. Red color indicated increased activity, while blue color indicated decreased activity in NCAM1 + EpCAM + cells derived organoids.

    Journal: Advanced Science

    Article Title: Expanded Hepatic Progenitor Cells Featured with Aggregation of α‐Synuclein Contribute to Pathologic Bile Duct Regeneration in Biliary Atresia

    doi: 10.1002/advs.76054

    Figure Lengend Snippet: Characterization of NCAM1 + EpCAM + HPCs in the expanded biliary clusters in BA. (a) Enrichment score of HPCs gene set in each biliary cluster. Comparisons among multiple clusters were performed using the Kruskal‐Wallis test, with a significance level of p < 0.001. (b) Venn diagram of DEGs showing higher expression in cluster 17 than other three biliary clusters (avg_log 2 FC>1), the DEGs showing higher expression in BA than the control group across all four biliary clusters (avg_log 2 FC>1), and the membrane protein database. (c) Gene density plot of NCAM1 in biliary clusters. (d): The proportion of NCAM1 positive cells of BA, Ctrl and external single‐cell dataset. P value was calculated by Chi‐squared test. (e): The proportion of NCAM1 + cells in EpCAM + cells of BA, Ctrl and normal cholangiocyte dataset ( GSE185477 contains four healthy human liver caudate lobes). P value was calculated by Chi‐squared test. (f): The expression of NCAM1 in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (g): The expression of NCAM1 in BA (n = 38) and Ctrl liver tissues ( n = 21). P value was calculated by unpaired t test. (h) Immunohistochemical staining of NCAM1 on paraffin sections of livers from Ctrl, BA with mild liver fibrosis (BA‐FM) and BA with severe liver fibrosis (BA‐FS). Scale bar: 100 µm. (i) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA ( n = 42) and Ctrl (n = 10). P value was calculated by unpaired t test. (j) Comparison of H‐scores for NCAM1 in portal areas of liver tissues between BA‐FM ( n = 30) and BA‐FS ( n = 12). P value was calculated by unpaired t test. (k) Morphology of NCAM1 + EpCAM + cells derived organoids and NCAM1 − EpCAM + cells derived organoids in BA (The left side of each group, scale bar: 200 µm). Regions highlighted with dotted boxes were magnified and shown on the right (scale bar: 50 µm). Calcein AM staining (Green) of the organoids were shown. (l) Volcano plot of DEGs of organoids derived from NCAM1 + EpCAM + cells ( n = 3) and NCAM1 − EpCAM + cells ( n = 4). (m) Bar chart showing transcriptional activity of TFs implicated in biliary and hepatic lineage. Red color indicated increased activity, while blue color indicated decreased activity in NCAM1 + EpCAM + cells derived organoids.

    Article Snippet: After single cells (viability>80%) were obtained, NCAM1 (CD56)‐positive and ‐negative expressing cells were isolated according to the REAlease CD56 MicroBead kit manual (130‐117‐033; Miltenyi, USA).

    Techniques: Expressing, Control, Membrane, Single Cell, Immunohistochemical staining, Staining, Comparison, Derivative Assay, Activity Assay

    NCAM1 + EpCAM + HPCs displayed transcriptomics associated with PD and α‐synuclein pathway. (a) Clinical variation analysis of the top 20 genes in cluster 17 calculated by Hypergeometric test. (b) Gene Set Variation Analysis of α‐synuclein pathway in each biliary cluster (cluster 7, 10, 15 and 17). The Kruskal‐Wallis test was used for multi‐group comparisons. (c) Gene Set Enrichment Analysis of α‐synuclein pathway in DEGs from organoids derived from NCAM1 + EpCAM + and NCAM1 − EpCAM + cells. p value was calculated by Permutation test. (d): The expression of α‐synuclein in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (e): The expression of α‐synuclein in BA ( n = 38) and Ctrl liver tissues ( n = 17). P value was calculated by unpaired t test. (f) Representative immunofluorescence images of NCAM1, EpCAM and α‐synuclein in BA and control sample. Scale bar: 20 µm. (g) Immunofluorescence images of Thioflavin‐S and α‐synuclein co‐staining in 3 patients with BA. The white triangle points to the co‐localization region. Scale bar: 50 µm. (h) Histogram showing serum total α‐synuclein in 40 BAs and 20 controls. p value was calculated by unpaired t test. (i) ROC of serum total α‐synuclein combined with or without GGT as a predictor for BA. (j) The mRNA expression of Snca and Ncam1 in RRV ( n = 7) and Control liver tissues ( n = 7) after log10 conversion. P value was calculated by unpaired t test. (k) Representative immunofluorescence images of NCAM1, EpCAM and α‐synuclein in RRV and control liver tissues. Scale bar: 100 µm. (l) Representative morphology and whole‐mount immunofluorescent staining of KRT19 and α‐synuclein in PFFs‐treated (PFFs) and untreated (Normal) mouse single cell derived CLC organoids. Scale bar of bright field images: 200 µm. Scale bar of immunofluorescent staining image: 100 µm. (m) Relative expression of hepatic and biliary markers in PFFs treated mouse single cell derived CLC organoids ( n = 3) and normal single cell derived CLC organoids ( n = 3). P value was calculated by unpaired t test. * p < 0.05; ** p < 0.01; *** p < 0.001.

    Journal: Advanced Science

    Article Title: Expanded Hepatic Progenitor Cells Featured with Aggregation of α‐Synuclein Contribute to Pathologic Bile Duct Regeneration in Biliary Atresia

    doi: 10.1002/advs.76054

    Figure Lengend Snippet: NCAM1 + EpCAM + HPCs displayed transcriptomics associated with PD and α‐synuclein pathway. (a) Clinical variation analysis of the top 20 genes in cluster 17 calculated by Hypergeometric test. (b) Gene Set Variation Analysis of α‐synuclein pathway in each biliary cluster (cluster 7, 10, 15 and 17). The Kruskal‐Wallis test was used for multi‐group comparisons. (c) Gene Set Enrichment Analysis of α‐synuclein pathway in DEGs from organoids derived from NCAM1 + EpCAM + and NCAM1 − EpCAM + cells. p value was calculated by Permutation test. (d): The expression of α‐synuclein in BA compared with normal liver tissues in GSE122340 (171 BA and 7 normal liver tissues) calculated by wilcoxon rank‐sum test. (e): The expression of α‐synuclein in BA ( n = 38) and Ctrl liver tissues ( n = 17). P value was calculated by unpaired t test. (f) Representative immunofluorescence images of NCAM1, EpCAM and α‐synuclein in BA and control sample. Scale bar: 20 µm. (g) Immunofluorescence images of Thioflavin‐S and α‐synuclein co‐staining in 3 patients with BA. The white triangle points to the co‐localization region. Scale bar: 50 µm. (h) Histogram showing serum total α‐synuclein in 40 BAs and 20 controls. p value was calculated by unpaired t test. (i) ROC of serum total α‐synuclein combined with or without GGT as a predictor for BA. (j) The mRNA expression of Snca and Ncam1 in RRV ( n = 7) and Control liver tissues ( n = 7) after log10 conversion. P value was calculated by unpaired t test. (k) Representative immunofluorescence images of NCAM1, EpCAM and α‐synuclein in RRV and control liver tissues. Scale bar: 100 µm. (l) Representative morphology and whole‐mount immunofluorescent staining of KRT19 and α‐synuclein in PFFs‐treated (PFFs) and untreated (Normal) mouse single cell derived CLC organoids. Scale bar of bright field images: 200 µm. Scale bar of immunofluorescent staining image: 100 µm. (m) Relative expression of hepatic and biliary markers in PFFs treated mouse single cell derived CLC organoids ( n = 3) and normal single cell derived CLC organoids ( n = 3). P value was calculated by unpaired t test. * p < 0.05; ** p < 0.01; *** p < 0.001.

    Article Snippet: After single cells (viability>80%) were obtained, NCAM1 (CD56)‐positive and ‐negative expressing cells were isolated according to the REAlease CD56 MicroBead kit manual (130‐117‐033; Miltenyi, USA).

    Techniques: Transcriptomics, Derivative Assay, Expressing, Immunofluorescence, Control, Staining, Single Cell