Structured Review

Becton Dickinson allophycocyanin apc conjugated anti cd34
Allophycocyanin Apc Conjugated Anti Cd34, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/allophycocyanin apc conjugated anti cd34/product/Becton Dickinson
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
allophycocyanin apc conjugated anti cd34 - by Bioz Stars, 2021-05
86/100 stars

Images

Related Articles

Incubation:

Article Title: Genomic safe harbors permit high ?-globin transgene expression in thalassemia induced pluripotent stem cells
Article Snippet: Undifferentiated iPS cells were dissociated with accutase, stained with Alexa Fluor 647-conjugated anti-Tra-1–81 or anti-Tra-1-60 or anti-SSEA3 or anti-SSEA4 and PE-Cy5-conjugated anti-HLA-ABC antibodies (BD Biosciences). .. The erythroid progeny of iPS cells were incubated with allophycocyanin (APC)-conjugated anti-CD34 or APC-conjugated anti-glycophorin A (GPA), PerCP-conjugated anti-CD45 and PE-conjugated anti-CD71 (BD Biosciences). .. Data were acquired in a LSRII cytometer (BD Biosciences) and analyzed with the FlowJo software (version 8.8.4; Tree Star).

Article Title: Molecular Profiling Reveals Similarities and Differences Between Primitive Subsets of Hematopoietic Cells Generated In Vitro from Human Embryonic Stem Cells and In Vivo during Embryogenesis
Article Snippet: A population enriched in CD34+ cells was isolated immediately after thawing the cells using the EasySep kit (Stem Cell Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions. .. The CD34+ -enriched cells were then incubated overnight in Hanks’ Balanced Salt Solution plus 50% fetal calf serum (FCS) at 4°C prior to being stained with allophycocyanin (APC)-conjugated anti-CD34 (8G12), phycoerythrin (PE)-conjugated anti-CD38 (HB7) from and fluorescein isothiocyanate (FITC)-conjugated anti-CD2, CD3, CD7, CD14, CD16, CD 19, CD20, CD24, CD36, CD45RA CD56, CD66b, CD71, and GlycophorinA (all antibodies from either BD Biosciences, San Jose, CA or StemCell Technologies). .. The CD38− (PE− ) and CD38+ (PE+ ) subsets of cells in the CD34+ (APC+ ) gate and negative for CD2, CD3, CD7, CD14, CD16, CD 19, CD20, CD24, CD36, CD45RA CD56, CD66b, CD71 and GlycophorinA (FITC− ) were collected at ≥95% purity by double sorting using a FACSVantage SE (BD Biosciences).

Article Title: Satellite Cells and Markers of Muscle Regeneration during Unloading and Reloading: Effects of Treatment with Resveratrol and Curcumin
Article Snippet: Cells were resuspended in 1 × 106 cell/100 µL FACS-specific buffer. .. All the cells were incubated with antibodies used to specifically identify the satellite cells: Phycoerythrin (PE)-conjugated anti-alpha-7 integrin (Ablab, Vancouver, Canada), a heterodimeric integral membrane protein critical for the modulation of cell–matrix interactions, and allophycocyanin (APC)-conjugated anti-CD34 (BD Pharmigen, San Jose, CA, USA), as a cell surface sialomucin (a mucopolysaccharide molecule containing sialic acid) with reported anti-adhesive, motile, and pre-proliferative properties for 30 min. Additionally, the cells from all study groups of mice were incubated at the same time (30 min) with specific antibodies to exclude other cell type populations that might also have been present in the muscle extracts, namely endothelial cells, leukocytes and hematopoietic cells: PE-cyanine7 (Cy7)-conjugated anti-CD31 (Biolegend, San Diego, CA, USA), a marker of endothelial cells, both anti-CD11b (Biolegend) and anti-CD45 (Biolegend) as markers of leukocytes, and anti-Sca-1 (Biolegend), a marker of hematopoietic stem cells. ..

Staining:

Article Title: Molecular Profiling Reveals Similarities and Differences Between Primitive Subsets of Hematopoietic Cells Generated In Vitro from Human Embryonic Stem Cells and In Vivo during Embryogenesis
Article Snippet: A population enriched in CD34+ cells was isolated immediately after thawing the cells using the EasySep kit (Stem Cell Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions. .. The CD34+ -enriched cells were then incubated overnight in Hanks’ Balanced Salt Solution plus 50% fetal calf serum (FCS) at 4°C prior to being stained with allophycocyanin (APC)-conjugated anti-CD34 (8G12), phycoerythrin (PE)-conjugated anti-CD38 (HB7) from and fluorescein isothiocyanate (FITC)-conjugated anti-CD2, CD3, CD7, CD14, CD16, CD 19, CD20, CD24, CD36, CD45RA CD56, CD66b, CD71, and GlycophorinA (all antibodies from either BD Biosciences, San Jose, CA or StemCell Technologies). .. The CD38− (PE− ) and CD38+ (PE+ ) subsets of cells in the CD34+ (APC+ ) gate and negative for CD2, CD3, CD7, CD14, CD16, CD 19, CD20, CD24, CD36, CD45RA CD56, CD66b, CD71 and GlycophorinA (FITC− ) were collected at ≥95% purity by double sorting using a FACSVantage SE (BD Biosciences).

Article Title: Enhanced Long-Term Transduction and Multilineage Engraftment of Human Hematopoietic Stem Cells Transduced with Tyrosine-Modified Recombinant Adeno-Associated Virus Serotype 2
Article Snippet: In vivo engraftment of human cells in both the bone marrow and spleen of xenografted mice was analyzed as described previously (Santat et al. , ). .. Lineage distribution was assessed in bone marrow and spleen cell suspensions after staining with human specific antibodies: FITC-conjugated anti-CD45; FITC- or allophycocyanin (APC)-conjugated anti-CD34; APC-conjugated anti-CD33, anti-CD14, and anti-glycophorin A; phycoerythrin (PE)-conjugated anti-CD19; and FITC-, PE- and APC-conjugated IgG controls (BD Biosciences, Mountain View, CA). .. Bone marrow lineages were sorted by fluorescence-activated cell sorting (FACS) using FITC–CD34, APC–CD33, PE–CD19, and APC–glycophorin A, as well as the appropriate controls.

Article Title: Metabolic Correction of Congenital Erythropoietic Porphyria with iPSCs Free of Reprogramming Factors
Article Snippet: .. Cells were stained with phycoerythrin (PE)- or allophycocyanin (APC)-conjugated anti-CD34, PECy5- or PE-conjugated anti-CD45, PE-conjugated anti-CD14, APC-conjugated anti-CD19, PE-conjugated anti-CD33, PE-conjugated anti-CD71, and APC-conjugated anti-GpA (all from BD, Franklin Lakes, NJ, USA). ..

Article Title: Gene Transfer Properties and Structural Modeling of Human Stem Cell-derived AAV
Article Snippet: In vivo engraftment of human cells in both the bone marrow and spleen of xenografted mice was analyzed as described previously. .. Lineage distribution was assessed in bone marrow and spleen cell suspensions following staining with human specific antibodies: FITC-conjugated anti-CD45, fluorescein isothiocyanate (FITC)- or allophycocyanin (APC)-conjugated anti-CD34, APC-conjugated anti-CD33 and anti-CD14, anti-Glycophorin A, phycoerythrin (PE)-conjugated anti-CD19, and FITC-, PE-, and APC-conjugated IgG controls (Becton Dickinson, Mountain View, CA). .. Bone marrow lineages were sorted by fluorescence-activated cell sorting (FACS) (Becton Dickinson) using FITC-CD34, APC-CD33, PE-CD19, and Glycophorin A-APC, as well as the appropriate controls.

Purification:

Article Title: How do megakaryocytic microparticles target and deliver cargo to alter the fate of hematopoietic stem cells?
Article Snippet: Size standard fluorescent beads (0.22, 0.45, 0.88 and 1.34 µm) and AccuCount fluorescent particles (~5.0 µm) were from SpheroTech. .. Fluorescein isothiocyanate (FITC) - or phycoerythrin (PE)-conjugated anti-CD41 (GPαIIb), PE-conjugated anti-CD62P (P-selectin), allophycocyanin (APC)-conjugated anti-CD34, PE-conjugated anti-CD11b, APC-conjugated anti-CD235a, FITC-conjugated CD63, APC-conjugated CD81 and purified anti-CD41, anti-CD42b, anti-CD43, anti-CD50 antibodies as well as corresponding IgG isotype were all from BD Bioscience. .. APC-conjugated anti-CD133 antibody was obtained from Miltenyi Biotec.

Mouse Assay:

Article Title: Satellite Cells and Markers of Muscle Regeneration during Unloading and Reloading: Effects of Treatment with Resveratrol and Curcumin
Article Snippet: Cells were resuspended in 1 × 106 cell/100 µL FACS-specific buffer. .. All the cells were incubated with antibodies used to specifically identify the satellite cells: Phycoerythrin (PE)-conjugated anti-alpha-7 integrin (Ablab, Vancouver, Canada), a heterodimeric integral membrane protein critical for the modulation of cell–matrix interactions, and allophycocyanin (APC)-conjugated anti-CD34 (BD Pharmigen, San Jose, CA, USA), as a cell surface sialomucin (a mucopolysaccharide molecule containing sialic acid) with reported anti-adhesive, motile, and pre-proliferative properties for 30 min. Additionally, the cells from all study groups of mice were incubated at the same time (30 min) with specific antibodies to exclude other cell type populations that might also have been present in the muscle extracts, namely endothelial cells, leukocytes and hematopoietic cells: PE-cyanine7 (Cy7)-conjugated anti-CD31 (Biolegend, San Diego, CA, USA), a marker of endothelial cells, both anti-CD11b (Biolegend) and anti-CD45 (Biolegend) as markers of leukocytes, and anti-Sca-1 (Biolegend), a marker of hematopoietic stem cells. ..

Marker:

Article Title: Satellite Cells and Markers of Muscle Regeneration during Unloading and Reloading: Effects of Treatment with Resveratrol and Curcumin
Article Snippet: Cells were resuspended in 1 × 106 cell/100 µL FACS-specific buffer. .. All the cells were incubated with antibodies used to specifically identify the satellite cells: Phycoerythrin (PE)-conjugated anti-alpha-7 integrin (Ablab, Vancouver, Canada), a heterodimeric integral membrane protein critical for the modulation of cell–matrix interactions, and allophycocyanin (APC)-conjugated anti-CD34 (BD Pharmigen, San Jose, CA, USA), as a cell surface sialomucin (a mucopolysaccharide molecule containing sialic acid) with reported anti-adhesive, motile, and pre-proliferative properties for 30 min. Additionally, the cells from all study groups of mice were incubated at the same time (30 min) with specific antibodies to exclude other cell type populations that might also have been present in the muscle extracts, namely endothelial cells, leukocytes and hematopoietic cells: PE-cyanine7 (Cy7)-conjugated anti-CD31 (Biolegend, San Diego, CA, USA), a marker of endothelial cells, both anti-CD11b (Biolegend) and anti-CD45 (Biolegend) as markers of leukocytes, and anti-Sca-1 (Biolegend), a marker of hematopoietic stem cells. ..

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Becton Dickinson apc mouse anti human cd34
    Mesenchymal stem cell (MSC) characterization in normoxic and hypoxic conditions. a Positive MSC markers (CD105, CD73, and CD90) and b negative MSC markers (CD,14, <t>CD34,</t> CD45)
    Apc Mouse Anti Human Cd34, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/apc mouse anti human cd34/product/Becton Dickinson
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    apc mouse anti human cd34 - by Bioz Stars, 2021-05
    95/100 stars
      Buy from Supplier

    86
    Becton Dickinson anti mouse sca 1
    Detection of endothelial/myo-endothelial cells (MECs) in adult mouse muscle (A–C) Co-localization of <t>Sca-1</t> (green), CD31 (red) and laminin (purple) shows co-expression of Sca-1 and CD31 in cells that reside between myofibers with (arrowheads) or without (arrows) their own basal lamina. Scale bar: 20 μm. (D, E) Immunostaining for laminin (purple) CD31 (green) Pax7 (red), and nuclei (DAPI) detects satellite cells, which reside beneath the basal lamina and are CD31 neg Pax7 pos (D, yellow arrow) as well as CD31 pos Pax7 neg cells (D, white arrowheads), which constitute the vast majority of CD31 pos cells, as well as rare CD31 pos Pax7 pos cells that reside within the interstitial space and are surrounded by laminin on both sides of the cell (E, white arrow). Scale bar: 20 μm.
    Anti Mouse Sca 1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mouse sca 1/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mouse sca 1 - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    Image Search Results


    Mesenchymal stem cell (MSC) characterization in normoxic and hypoxic conditions. a Positive MSC markers (CD105, CD73, and CD90) and b negative MSC markers (CD,14, CD34, CD45)

    Journal: Stem Cell Research & Therapy

    Article Title: Hypoxic preconditioning induces epigenetic changes and modifies swine mesenchymal stem cell angiogenesis and senescence in experimental atherosclerotic renal artery stenosis

    doi: 10.1186/s13287-021-02310-z

    Figure Lengend Snippet: Mesenchymal stem cell (MSC) characterization in normoxic and hypoxic conditions. a Positive MSC markers (CD105, CD73, and CD90) and b negative MSC markers (CD,14, CD34, CD45)

    Article Snippet: Conversely, MSC were expected to not express CD45 (Abcam, Cat. # ab51482), CD34 (BD Biosciences, San Jose, CA, Cat. # 340441), or CD14 (Abcam, Cat. # ab82012).

    Techniques:

    Phenotypic diversity of acute myeloid leukemia (AML) leukemic stem cells (LSCs). (A) CD34 and CD38 surface marker profiles of all 52 patients. (B) Gating strategy for AML-LSCs. (C) Three distinct populations were analyzed as LSC phenotypes: left, multipotent progenitor (MPP)-like LSC; middle, lymphoid primed multipotent progenitor (LMPP)-like LSC; right, granulocyte-macrophage progenitors (GMP)-like LSC.

    Journal: The Korean Journal of Internal Medicine

    Article Title: Leukemic stem cell phenotype is associated with mutational profile in acute myeloid leukemia

    doi: 10.3904/kjim.2020.014

    Figure Lengend Snippet: Phenotypic diversity of acute myeloid leukemia (AML) leukemic stem cells (LSCs). (A) CD34 and CD38 surface marker profiles of all 52 patients. (B) Gating strategy for AML-LSCs. (C) Three distinct populations were analyzed as LSC phenotypes: left, multipotent progenitor (MPP)-like LSC; middle, lymphoid primed multipotent progenitor (LMPP)-like LSC; right, granulocyte-macrophage progenitors (GMP)-like LSC.

    Article Snippet: Cells were stained with following anti-human monoclonal antibodies: CD45-APC/cy7 (557833), CD34-APC (555824), CD38-BV421 (562445), CD90-PE (555596), CD123-PE/Cy7 (560826), and CD45RA-PerCP/Cy5.5 (563429) (BD Bioscience).

    Techniques: Marker

    A comparison of the gene expression profiles of single-purified human CB-derived CD34 + and CD34 − and CD90 + HSCs by qRT-PCR. Multi-plex (79 genes) single-cell qRT-PCR using CD34 + and CD34 − HSCs and CD90 + HSCs was performed. a The 54 gene expression profiles in CD34 + and CD34 − HSCs and CD90 + HSC are depicted using a heatmap. These 54 genes were classified as HSC/HPC-, erythrocyte-, megakaryocyte-, myeloid cell-, lymphocyte- and epigenetics-related genes. b The expression of individual genes in CD34 + and CD34 − HSCs and CD90 + HSCs is depicted by violin plots. The HSC maintenance genes highly expressed in both CD34 + and CD34 − HSCs and CD90 + HSCs ( KIT , RUNK1 , TAL1 , BMI1 , DNMT3A , TGFBR1 and R2 ) are shown in the upper panel and highlighted by green color in a . The genes highly expressed in CD34 + HSCs ( IFITM1 , MPL , IKZF1 , ETV6 , ALDH1A1 and IGF1R ) are shown in the middle left panel and highlighted by pink color in a . The genes highly expressed in CD34 − HSCs ( EZH2 and MYB ) are shown in the middle right panel and highlighted by blue color in a . The gate names (R6 and R8) presented in this figure correspond to the same fractions in Fig. 1e, f . c Violin plots of the reference genes, including ACTB , GAPDH , PTPRC (CD45) and PGK-1 (left). Violin plots of the genes for CD34 and PROM1 (CD133) (middle). qRT-PCR of the VNN2 (GPI-80) mRNA expression in the 18Lin − CD34 + CD38 − CD133 + GPI-80 +/ − and 18Lin − CD34 − CD133 + GPI-80 +/ − cells, compared with the positive control (CB-derived neutrophils) and negative control (THP-1 cells) (right). n.d.: not detected

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: A comparison of the gene expression profiles of single-purified human CB-derived CD34 + and CD34 − and CD90 + HSCs by qRT-PCR. Multi-plex (79 genes) single-cell qRT-PCR using CD34 + and CD34 − HSCs and CD90 + HSCs was performed. a The 54 gene expression profiles in CD34 + and CD34 − HSCs and CD90 + HSC are depicted using a heatmap. These 54 genes were classified as HSC/HPC-, erythrocyte-, megakaryocyte-, myeloid cell-, lymphocyte- and epigenetics-related genes. b The expression of individual genes in CD34 + and CD34 − HSCs and CD90 + HSCs is depicted by violin plots. The HSC maintenance genes highly expressed in both CD34 + and CD34 − HSCs and CD90 + HSCs ( KIT , RUNK1 , TAL1 , BMI1 , DNMT3A , TGFBR1 and R2 ) are shown in the upper panel and highlighted by green color in a . The genes highly expressed in CD34 + HSCs ( IFITM1 , MPL , IKZF1 , ETV6 , ALDH1A1 and IGF1R ) are shown in the middle left panel and highlighted by pink color in a . The genes highly expressed in CD34 − HSCs ( EZH2 and MYB ) are shown in the middle right panel and highlighted by blue color in a . The gate names (R6 and R8) presented in this figure correspond to the same fractions in Fig. 1e, f . c Violin plots of the reference genes, including ACTB , GAPDH , PTPRC (CD45) and PGK-1 (left). Violin plots of the genes for CD34 and PROM1 (CD133) (middle). qRT-PCR of the VNN2 (GPI-80) mRNA expression in the 18Lin − CD34 + CD38 − CD133 + GPI-80 +/ − and 18Lin − CD34 − CD133 + GPI-80 +/ − cells, compared with the positive control (CB-derived neutrophils) and negative control (THP-1 cells) (right). n.d.: not detected

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: Expressing, Purification, Derivative Assay, Quantitative RT-PCR, Positive Control, Negative Control

    A comparison of the gene expression profiles between CD34 + and CD34 − SRCs (HSCs) by a microarray. a The gene expression profiles of CD34 + and CD34 − SRCs (HSCs) were compared by a Gene Set Enrichment Analysis (GSEA) using the data from the microarray analysis. The gene sets enriched in CD34 − SRCs (blue bar) and those in CD34 + SRCs (red bar) with a Normalized Enrichment Score

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: A comparison of the gene expression profiles between CD34 + and CD34 − SRCs (HSCs) by a microarray. a The gene expression profiles of CD34 + and CD34 − SRCs (HSCs) were compared by a Gene Set Enrichment Analysis (GSEA) using the data from the microarray analysis. The gene sets enriched in CD34 − SRCs (blue bar) and those in CD34 + SRCs (red bar) with a Normalized Enrichment Score

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: Expressing, Microarray

    The current and proposed models for human HSC hierarchy. a The current model 1 – 3 for the human HSC hierarchy. CD34 + HSCs as defined by a CD34 + CD38 − CD45RA − CD90 + CD49f + immunophenotype differentiate into MPPs, CMPs, MLPs, GMPs and MEPs. b The model 19 , 20 proposed based on our series of studies 18 , 21 , 22 , 25 , 33 – 36 , 60 , in which CD34 − HSCs are defined by a CD34 − CD38 low/ − CD45RA − FLT3 − CD110 − CD133 + GPI-80 + immunophenotype. CD34 − HSCs generate CD34 + HSCs in vitro 25 , 35 , 36 and in vivo 18 , 33 , suggesting that CD34 − HSCs reside at the apex of human HSC hierarchy. CD34 − HSCs, then, differentiate into MPPs, CMPs, GMPs and MEPs according to the current model 1 – 3 . Incorporating the present studies, a revised road map, which allows a commitment/differentiation pathway of CD34 − HSCs directly into MEPs (bypass route), is shown. MPP: multipotent progenitor, MLP: multilymphoid progenitor, CMP: common myeloid progenitor, GMP: granulocyte/macrophage progenitor, MEP: megakaryocyte/erythrocyte progenitor

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: The current and proposed models for human HSC hierarchy. a The current model 1 – 3 for the human HSC hierarchy. CD34 + HSCs as defined by a CD34 + CD38 − CD45RA − CD90 + CD49f + immunophenotype differentiate into MPPs, CMPs, MLPs, GMPs and MEPs. b The model 19 , 20 proposed based on our series of studies 18 , 21 , 22 , 25 , 33 – 36 , 60 , in which CD34 − HSCs are defined by a CD34 − CD38 low/ − CD45RA − FLT3 − CD110 − CD133 + GPI-80 + immunophenotype. CD34 − HSCs generate CD34 + HSCs in vitro 25 , 35 , 36 and in vivo 18 , 33 , suggesting that CD34 − HSCs reside at the apex of human HSC hierarchy. CD34 − HSCs, then, differentiate into MPPs, CMPs, GMPs and MEPs according to the current model 1 – 3 . Incorporating the present studies, a revised road map, which allows a commitment/differentiation pathway of CD34 − HSCs directly into MEPs (bypass route), is shown. MPP: multipotent progenitor, MLP: multilymphoid progenitor, CMP: common myeloid progenitor, GMP: granulocyte/macrophage progenitor, MEP: megakaryocyte/erythrocyte progenitor

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: In Vitro, In Vivo

    Representative FACS profile and colony-forming capacity of highly purified CB-derived 18Lin - CD34 + CD38 - CD133 + GPI-80 +/ − and 18Lin - CD34 − CD133 + GPI-80 +/ − cells. A representative FACS profile is shown. a The forward scatter/side scatter (FSC/SSC) profile of immunomagnetically separated Lin − cells. The R1 gate was set on the blast-lymphocyte window. b The R2 gate was set on the 18Lin − living cells. c The R2 gated cells were subdivided into two fractions: 18Lin − CD45 + CD34 + (R3) and CD34 − (R4) cells, according to their expression of CD34. The definitions of CD34 +/ − cells are as follows: the CD34 + fraction contains cells expressing > 5% of the maximum BV421 fluorescence intensity (FI). The CD34 − level of FI was determined based on the Fluorescence Minus One controls. d The cells residing in the R3 gate were further subdivided into 18Lin − CD45 + CD34 + CD38 − (R5) cells. The CD38 − fraction contains cells expressing

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: Representative FACS profile and colony-forming capacity of highly purified CB-derived 18Lin - CD34 + CD38 - CD133 + GPI-80 +/ − and 18Lin - CD34 − CD133 + GPI-80 +/ − cells. A representative FACS profile is shown. a The forward scatter/side scatter (FSC/SSC) profile of immunomagnetically separated Lin − cells. The R1 gate was set on the blast-lymphocyte window. b The R2 gate was set on the 18Lin − living cells. c The R2 gated cells were subdivided into two fractions: 18Lin − CD45 + CD34 + (R3) and CD34 − (R4) cells, according to their expression of CD34. The definitions of CD34 +/ − cells are as follows: the CD34 + fraction contains cells expressing > 5% of the maximum BV421 fluorescence intensity (FI). The CD34 − level of FI was determined based on the Fluorescence Minus One controls. d The cells residing in the R3 gate were further subdivided into 18Lin − CD45 + CD34 + CD38 − (R5) cells. The CD38 − fraction contains cells expressing

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: FACS, Purification, Derivative Assay, Expressing, Fluorescence

    SCID-repopulating cell activity and serial analyses of human cell repopulation of 18Lin − CD34 + CD38 − CD133 + GPI-80 + and 18Lin − CD34 − CD133 + GPI-80 + cells in primary and secondary NOG mice by IBMI. a A schematic illustration of primary and secondary transplantation of CD34 + and CD34 − SRCs is shown. PR, primary recipient; SR, secondary recipient. b The long-term repopulating potential of the CD34 + and CD34 − SRCs was determined by serially analyzing the kinetics of BM engraftment for 20 to 22 weeks in primary NOG mice that received transplants of 200 18Lin − CD34 + CD38 − CD133 + GPI-80 + cells (41 SRCs) ( n = 25) and 200 18Lin − CD34 − CD133 + GPI-80 + cells (25 SRCs) ( n = 23), respectively. All of the primary recipient mice were highly repopulated with human CD45 + cells. The human CD45 + cell repopulation rates of the mice that were transplanted with CD34 + SRCs at 12 and 18 weeks after transplantation were significantly higher than those of the mice that were transplanted with CD34 − SRCs. However, the repopulation rates of both mice were comparable at 20 to 22 weeks after transplantation (mean ± S.D., * p

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: SCID-repopulating cell activity and serial analyses of human cell repopulation of 18Lin − CD34 + CD38 − CD133 + GPI-80 + and 18Lin − CD34 − CD133 + GPI-80 + cells in primary and secondary NOG mice by IBMI. a A schematic illustration of primary and secondary transplantation of CD34 + and CD34 − SRCs is shown. PR, primary recipient; SR, secondary recipient. b The long-term repopulating potential of the CD34 + and CD34 − SRCs was determined by serially analyzing the kinetics of BM engraftment for 20 to 22 weeks in primary NOG mice that received transplants of 200 18Lin − CD34 + CD38 − CD133 + GPI-80 + cells (41 SRCs) ( n = 25) and 200 18Lin − CD34 − CD133 + GPI-80 + cells (25 SRCs) ( n = 23), respectively. All of the primary recipient mice were highly repopulated with human CD45 + cells. The human CD45 + cell repopulation rates of the mice that were transplanted with CD34 + SRCs at 12 and 18 weeks after transplantation were significantly higher than those of the mice that were transplanted with CD34 − SRCs. However, the repopulation rates of both mice were comparable at 20 to 22 weeks after transplantation (mean ± S.D., * p

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: Activity Assay, Mouse Assay, Transplantation Assay

    Single-cell-based gene expression profiles of human CB-derived CD34 + and CD34 − HSCs. Using highly purified human CB-derived 18Lin − CD34 + CD38 − CD133 + GPI-80 + and 18Lin − CD34 − CD133 + GPI-80 + cells, we performed a single-cell-based gene expression analysis. The immunophenotypes of target cells, including controls, are presented in Supplementary Data 6 and the 79 target genes that play important roles in the pathway of HSC development/differentiation 63 are listed in Supplementary Data 7 . a A principal component analysis (PCA) revealed that the gene expression profiles in individual CD34 + ( n = 33) and CD34 − HSCs ( n = 23) were clearly different. The gene expression profiles of individual CD90 + HSCs ( n = 5) were similar to those of CD34 + HSCs but not to those of CD34 − HSCs. The dotted line represents the border region between the CD34 + and CD34 − HSCs calculated by a Fisher’s linear discriminant analysis. b An unsupervised hierarchical clustering analysis (Dendrogram) clearly showed two clusters, 1 and 2. Interestingly, all CD34 − HSCs belong to cluster 1. Three subgroups were detected in cluster 1. The left-most subgroup uniformly contained 10 CD34 − HSCs. The remaining 13 CD34 − HSCs were scattered between the other two subgroups mixed with CD34 + HSCs and CD90 + HSCs and MPPs. In contrast, some of the CD34 + HSCs and CD90 + HSCs, most of the MPP, and all other HPCs (CMP, GMP, MEP and MLP) belonged to cluster 2. These results demonstrated that the gene expression profiles of CD34 − HSC were unique and largely differed from those of other classes of CD34 + HSPCs. c A hierarchical clustering analysis of 62 genes (heatmap) detected 3 clusters ( a – c ), which are highlighted with yellow squares. Cluster (A) contained HSC/HPC-related genes, including RUNX1 , BMI1 , DNMT3a and SCL/TAL1 , in addition to CD34 and PROM1 (CD133). Cluster (B) contained HPC-related genes, including MYB , FOXO3A , SMAD4 , NFE2 and NOTCH1 . Cluster (C) contained mature cell-related genes, including those for various cytokine receptors ( CSF2RA , IL-6R , CSF1R , IGF2R and IL-3RA )

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: Single-cell-based gene expression profiles of human CB-derived CD34 + and CD34 − HSCs. Using highly purified human CB-derived 18Lin − CD34 + CD38 − CD133 + GPI-80 + and 18Lin − CD34 − CD133 + GPI-80 + cells, we performed a single-cell-based gene expression analysis. The immunophenotypes of target cells, including controls, are presented in Supplementary Data 6 and the 79 target genes that play important roles in the pathway of HSC development/differentiation 63 are listed in Supplementary Data 7 . a A principal component analysis (PCA) revealed that the gene expression profiles in individual CD34 + ( n = 33) and CD34 − HSCs ( n = 23) were clearly different. The gene expression profiles of individual CD90 + HSCs ( n = 5) were similar to those of CD34 + HSCs but not to those of CD34 − HSCs. The dotted line represents the border region between the CD34 + and CD34 − HSCs calculated by a Fisher’s linear discriminant analysis. b An unsupervised hierarchical clustering analysis (Dendrogram) clearly showed two clusters, 1 and 2. Interestingly, all CD34 − HSCs belong to cluster 1. Three subgroups were detected in cluster 1. The left-most subgroup uniformly contained 10 CD34 − HSCs. The remaining 13 CD34 − HSCs were scattered between the other two subgroups mixed with CD34 + HSCs and CD90 + HSCs and MPPs. In contrast, some of the CD34 + HSCs and CD90 + HSCs, most of the MPP, and all other HPCs (CMP, GMP, MEP and MLP) belonged to cluster 2. These results demonstrated that the gene expression profiles of CD34 − HSC were unique and largely differed from those of other classes of CD34 + HSPCs. c A hierarchical clustering analysis of 62 genes (heatmap) detected 3 clusters ( a – c ), which are highlighted with yellow squares. Cluster (A) contained HSC/HPC-related genes, including RUNX1 , BMI1 , DNMT3a and SCL/TAL1 , in addition to CD34 and PROM1 (CD133). Cluster (B) contained HPC-related genes, including MYB , FOXO3A , SMAD4 , NFE2 and NOTCH1 . Cluster (C) contained mature cell-related genes, including those for various cytokine receptors ( CSF2RA , IL-6R , CSF1R , IGF2R and IL-3RA )

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: Expressing, Derivative Assay, Purification

    Human multi-lineage hematopoietic repopulation abilities of single CD34 + and CD34 − SRCs. Human multi-lineage hematopoietic repopulations in the primary recipient NSG mice that received a single 18Lin − CD34 + CD38 − CD133 + GPI-80 + (Mouse ID: 34 + NSG001) or b single 18Lin − CD34 − CD133 + GPI-80 + (Mouse ID: 34 − NSG027) cells were analyzed by 6-color FCM at 22 or 21 weeks after transplantation. The expression of CD19, CD33 CD34 (BM, PB and spleen), CD11b and CD14 (BM), CD41 (BM and spleen), CD235a (BM) and CD56 (spleen) in living human CD45 + cells was analyzed. The presence of CD41 + and CD235a + cells in BM was analyzed using a human CD45 +/ − cell gate (depicted by red dotted lines). The mouse ID numbers presented in the figure corresponded those listed in Supplementary Data 3a . Analyses of the BM (other bones), PB and spleens of the two representative mice that received either CD34 + SRC a or CD34 − SRC b revealed that both SRCs had an in vivo differentiation capacity comparable to that of CD34 + stem/progenitor cells, CD19 + B-lymphoids, CD33 + /CD11b + myeloids, CD14 + monocytes, CD235a + erythroid and CD41 + megakaryocytic lineages. CD56 + NK cells were detected in the spleen. These results confirmed that both CD34 + and CD34 − SRCs had definite multi-lineage differentiation potential. Detailed FCM data of all recipient mice that received single CD34 + and CD34 − SRCs are presented in Supplementary Data 3b

    Journal: Nature Communications

    Article Title: A revised road map for the commitment of human cord blood CD34-negative hematopoietic stem cells

    doi: 10.1038/s41467-018-04441-z

    Figure Lengend Snippet: Human multi-lineage hematopoietic repopulation abilities of single CD34 + and CD34 − SRCs. Human multi-lineage hematopoietic repopulations in the primary recipient NSG mice that received a single 18Lin − CD34 + CD38 − CD133 + GPI-80 + (Mouse ID: 34 + NSG001) or b single 18Lin − CD34 − CD133 + GPI-80 + (Mouse ID: 34 − NSG027) cells were analyzed by 6-color FCM at 22 or 21 weeks after transplantation. The expression of CD19, CD33 CD34 (BM, PB and spleen), CD11b and CD14 (BM), CD41 (BM and spleen), CD235a (BM) and CD56 (spleen) in living human CD45 + cells was analyzed. The presence of CD41 + and CD235a + cells in BM was analyzed using a human CD45 +/ − cell gate (depicted by red dotted lines). The mouse ID numbers presented in the figure corresponded those listed in Supplementary Data 3a . Analyses of the BM (other bones), PB and spleens of the two representative mice that received either CD34 + SRC a or CD34 − SRC b revealed that both SRCs had an in vivo differentiation capacity comparable to that of CD34 + stem/progenitor cells, CD19 + B-lymphoids, CD33 + /CD11b + myeloids, CD14 + monocytes, CD235a + erythroid and CD41 + megakaryocytic lineages. CD56 + NK cells were detected in the spleen. These results confirmed that both CD34 + and CD34 − SRCs had definite multi-lineage differentiation potential. Detailed FCM data of all recipient mice that received single CD34 + and CD34 − SRCs are presented in Supplementary Data 3b

    Article Snippet: The aliquots of BM aspirates were stained with a Pacific Blue (PB)-conjugated anti-human CD45 mAb (BioLegend), a PE-Cy7-conjugated anti-mouse CD45.1 mAb (Beckman Coulter), FITC-conjugated anti-human CD19 mAb, PE-conjugated anti-human CD33 mAb (Beckman Coulter), APC-conjugated anti-human CD34 mAb (BD Biosciences) and 7-AAD and were analyzed by six-color FCM (FACS CantoII).

    Techniques: Mouse Assay, Transplantation Assay, Expressing, In Vivo

    Detection of endothelial/myo-endothelial cells (MECs) in adult mouse muscle (A–C) Co-localization of Sca-1 (green), CD31 (red) and laminin (purple) shows co-expression of Sca-1 and CD31 in cells that reside between myofibers with (arrowheads) or without (arrows) their own basal lamina. Scale bar: 20 μm. (D, E) Immunostaining for laminin (purple) CD31 (green) Pax7 (red), and nuclei (DAPI) detects satellite cells, which reside beneath the basal lamina and are CD31 neg Pax7 pos (D, yellow arrow) as well as CD31 pos Pax7 neg cells (D, white arrowheads), which constitute the vast majority of CD31 pos cells, as well as rare CD31 pos Pax7 pos cells that reside within the interstitial space and are surrounded by laminin on both sides of the cell (E, white arrow). Scale bar: 20 μm.

    Journal: Nature communications

    Article Title: Intramuscular adipogenesis is inhibited by myo-endothelial progenitors with functioning Bmpr1a signaling

    doi: 10.1038/ncomms5063

    Figure Lengend Snippet: Detection of endothelial/myo-endothelial cells (MECs) in adult mouse muscle (A–C) Co-localization of Sca-1 (green), CD31 (red) and laminin (purple) shows co-expression of Sca-1 and CD31 in cells that reside between myofibers with (arrowheads) or without (arrows) their own basal lamina. Scale bar: 20 μm. (D, E) Immunostaining for laminin (purple) CD31 (green) Pax7 (red), and nuclei (DAPI) detects satellite cells, which reside beneath the basal lamina and are CD31 neg Pax7 pos (D, yellow arrow) as well as CD31 pos Pax7 neg cells (D, white arrowheads), which constitute the vast majority of CD31 pos cells, as well as rare CD31 pos Pax7 pos cells that reside within the interstitial space and are surrounded by laminin on both sides of the cell (E, white arrow). Scale bar: 20 μm.

    Article Snippet: Antibodies were added at the following dilutions: anti-mouse CD45 (1:300, PE-Cy7 conjugate, clone 30-F11), anti-mouse Sca-1 (Ly-6A/E, 1:100, FITC or APC-Cy7 conjugate, clone E13-161.7), anti-mouse CD31 (1:200, APC or PE conjugate, clone MEC13.3) (all from BD Pharmingen, CA), anti-mouse PDGFR1α (1:100, APC or biotin conjugate, clone APA5, Biolegend, CA), anti-mouse CD106 (VCAM-1, 1:75, FITC or PE conjugate, clone 429, Biolegend, CA, anti-mouse VE-cadherin (1:100, clone 11D4.1, BD Pharmingen, CA).

    Techniques: Expressing, Immunostaining