cd34 expression Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • cd34  (Abcam)
    99
    Abcam cd34
    CRC-secreted miR-25-3p primes pre-metastatic niche. a Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular permeability of mice liver by in vivo permeability assay. The mice were injected with rhodamine–dextran after exposure to PKH67-labeled exosomes. Levels of rhodamine–dextran fluorescence in tissues were quantified using Image J software and normalized to the levels of DAPI. Mean ± SEM are provided ( n = 5). Scale bar represents 50 µm. b Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular KLF4, KLF2, and ZO-1 expression (red) in hepatic vessels by immunofluorescence. The vascular structures were labeled by <t>CD34</t> (green). Scale bar represents 50 µm. c Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on KLF4, ZO-1, occludin, Claudin5, KLF2, VEGFR2, fibronectin, and S100 expression in mice liver by Western blot. d The mice were intra-spleen injected with naked SW480 cells after exposure to NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments. The number of liver metastatic sites (indicated by arrows) was counted under the microscope. Mean ± SEM are provided ( n = 5). Scale bar in left panels represents 0.5 cm. Scale bar in right panels represents 100 µm. ** P
    Cd34, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd34/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd34 - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    86
    Becton Dickinson cd34
    Long-Term Repopulation Patterns of <t>CD34+/–</t> MPL+/– SRCs in NOG Mice and Their Multilineage Differentiation Potentials
    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/cd34/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd34 - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    86
    Santa Cruz Biotechnology cd34
    Characterization of rat bone marrow-derived mesenchymal stromal cells (BMSCs). (a) The plastic-adherent cells demonstrated a homogeneous fibroblast-like and spindle-shaped morphology. Original magnification, ×100. (b) Adipogenic differentiation of BMSCs stained with oil red O. Original magnification, ×200. (c) Osteogenic differentiation of BM-MSCs stained with alizarin red. Original magnification, ×400. (d) FACS analysis demonstrated expression of markers attributed to BMSCs. The cells were devoid of hematopoietic cells as indicated by the lack of CD45 and <t>CD34.</t> The MSC-specific markers, CD29, CD44, CD73, CD105, and CD90 were strongly expressed on the cells.
    Cd34, supplied by Santa Cruz Biotechnology, 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/cd34/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd34 - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    86
    miltenyi biotec cd34
    Hypoxia reduces proliferation of <t>CD34</t> + hematopoietic stem and progenitor cells. CD34 + and CD133 + cells in cultures in both hypoxia and normoxia were determined by flow cytometry on days 1, 7, 14 and 21. (A) Fold change in the number of CD34 + cells in the normoxic and hypoxic cultures was calculated relative to day 1. (B) Fold change in the number of CD133 + cells in the normoxic and hypoxic cultures was calculated relative to day 1. (C) Percentage of CD34 + cells in the Lin − /Live population is shown. Data are represented as the mean with standard error ( n = 6 for CD34 and n = 4 for CD133). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p
    Cd34, supplied by miltenyi biotec, 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/cd34/product/miltenyi biotec
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd34 - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    N/A
    CD34 HEK293T cell transient overexpression lysate as WB positive control
      Buy from Supplier

    N/A
    CD34 HEK293T cell transient overexpression lysate as WB positive control
      Buy from Supplier

    N/A
    Full length Clone DNA of Human CD34 molecule transcript variant 1
      Buy from Supplier

    Image Search Results


    CRC-secreted miR-25-3p primes pre-metastatic niche. a Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular permeability of mice liver by in vivo permeability assay. The mice were injected with rhodamine–dextran after exposure to PKH67-labeled exosomes. Levels of rhodamine–dextran fluorescence in tissues were quantified using Image J software and normalized to the levels of DAPI. Mean ± SEM are provided ( n = 5). Scale bar represents 50 µm. b Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular KLF4, KLF2, and ZO-1 expression (red) in hepatic vessels by immunofluorescence. The vascular structures were labeled by CD34 (green). Scale bar represents 50 µm. c Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on KLF4, ZO-1, occludin, Claudin5, KLF2, VEGFR2, fibronectin, and S100 expression in mice liver by Western blot. d The mice were intra-spleen injected with naked SW480 cells after exposure to NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments. The number of liver metastatic sites (indicated by arrows) was counted under the microscope. Mean ± SEM are provided ( n = 5). Scale bar in left panels represents 0.5 cm. Scale bar in right panels represents 100 µm. ** P

    Journal: Nature Communications

    Article Title: Cancer-derived exosomal miR-25-3p promotes pre-metastatic niche formation by inducing vascular permeability and angiogenesis

    doi: 10.1038/s41467-018-07810-w

    Figure Lengend Snippet: CRC-secreted miR-25-3p primes pre-metastatic niche. a Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular permeability of mice liver by in vivo permeability assay. The mice were injected with rhodamine–dextran after exposure to PKH67-labeled exosomes. Levels of rhodamine–dextran fluorescence in tissues were quantified using Image J software and normalized to the levels of DAPI. Mean ± SEM are provided ( n = 5). Scale bar represents 50 µm. b Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular KLF4, KLF2, and ZO-1 expression (red) in hepatic vessels by immunofluorescence. The vascular structures were labeled by CD34 (green). Scale bar represents 50 µm. c Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on KLF4, ZO-1, occludin, Claudin5, KLF2, VEGFR2, fibronectin, and S100 expression in mice liver by Western blot. d The mice were intra-spleen injected with naked SW480 cells after exposure to NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments. The number of liver metastatic sites (indicated by arrows) was counted under the microscope. Mean ± SEM are provided ( n = 5). Scale bar in left panels represents 0.5 cm. Scale bar in right panels represents 100 µm. ** P

    Article Snippet: The following primary antibodies were used: CD34 (Abcam, ab81289, 1:200 dilution), KLF2 (Abcam, ab203591, 1:100 dilution), KLF4 (Abcam, ab106629, 1:100 dilution), HIFα (Affinity, AF1009, 1:200 dilution).

    Techniques: Permeability, Mouse Assay, In Vivo, Injection, Labeling, Fluorescence, Software, Expressing, Immunofluorescence, Western Blot, Microscopy

    CRC-secreted miR-25-3p is transferred to endothelial cells. a RT-PCR analysis of miR-25-3p expression in 27 paired fresh CRC tissues and matched adjacent normal mucosa. The expression of miR-25-3p in normal mucosa was normalized to 1. Mean ± SEM are provided ( n = 3). b RT-PCR analysis of miR-25-3p expression in 27 cases of the primary CRC tissues with or without metastasis (17 cases without metastasis, 10 cases with metastasis). Mean ± SEM are provided. c Correlation analysis of miR-25-3p (red) expression in colorectal epithelial cells/cancer cells (labeled by CK, purple) and their adjacent endothelial cells (labeled by CD34, green). MiR-25-3p levels were determined by FISH and scored as described in Methods. Pearson’s correlation coefficient (r) and P- value are shown ( n = 20). P- value is from Spearman’s test. Scale bar represents 50 µm. d Transmission electron microscopy of exosomes derived from SW480/mock and SW480/miR-25-3p. Scale bar represents 50 nm. e Western blotting analysis of TSG101 and CD63 in SW480/mock, SW480/miR-25-3p and their exosomes. f The top panels show presence of Cy3 fluorescence and PKH67 lipid dye in HUVECs after adding PKH67-labeled exosomes derived from SW480 cells for 48 h. HUVECs incubated with naked-miR-25-3p-Cy3 were used as a negative control (the bottom panels). Scale bar represent 10 µm. g RT-PCR analysis of miR-25-3p expression in HUVECs incubated with exosomes derived from SW480/mock and SW480/miR-25-3p for 3 h, 6 h, 12 h, 24 h, and 48 h. Mean ± SEM are provided ( n = 3). ** P

    Journal: Nature Communications

    Article Title: Cancer-derived exosomal miR-25-3p promotes pre-metastatic niche formation by inducing vascular permeability and angiogenesis

    doi: 10.1038/s41467-018-07810-w

    Figure Lengend Snippet: CRC-secreted miR-25-3p is transferred to endothelial cells. a RT-PCR analysis of miR-25-3p expression in 27 paired fresh CRC tissues and matched adjacent normal mucosa. The expression of miR-25-3p in normal mucosa was normalized to 1. Mean ± SEM are provided ( n = 3). b RT-PCR analysis of miR-25-3p expression in 27 cases of the primary CRC tissues with or without metastasis (17 cases without metastasis, 10 cases with metastasis). Mean ± SEM are provided. c Correlation analysis of miR-25-3p (red) expression in colorectal epithelial cells/cancer cells (labeled by CK, purple) and their adjacent endothelial cells (labeled by CD34, green). MiR-25-3p levels were determined by FISH and scored as described in Methods. Pearson’s correlation coefficient (r) and P- value are shown ( n = 20). P- value is from Spearman’s test. Scale bar represents 50 µm. d Transmission electron microscopy of exosomes derived from SW480/mock and SW480/miR-25-3p. Scale bar represents 50 nm. e Western blotting analysis of TSG101 and CD63 in SW480/mock, SW480/miR-25-3p and their exosomes. f The top panels show presence of Cy3 fluorescence and PKH67 lipid dye in HUVECs after adding PKH67-labeled exosomes derived from SW480 cells for 48 h. HUVECs incubated with naked-miR-25-3p-Cy3 were used as a negative control (the bottom panels). Scale bar represent 10 µm. g RT-PCR analysis of miR-25-3p expression in HUVECs incubated with exosomes derived from SW480/mock and SW480/miR-25-3p for 3 h, 6 h, 12 h, 24 h, and 48 h. Mean ± SEM are provided ( n = 3). ** P

    Article Snippet: The following primary antibodies were used: CD34 (Abcam, ab81289, 1:200 dilution), KLF2 (Abcam, ab203591, 1:100 dilution), KLF4 (Abcam, ab106629, 1:100 dilution), HIFα (Affinity, AF1009, 1:200 dilution).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Labeling, Fluorescence In Situ Hybridization, Transmission Assay, Electron Microscopy, Derivative Assay, Western Blot, Fluorescence, Incubation, Negative Control

    Long-Term Repopulation Patterns of CD34+/– MPL+/– SRCs in NOG Mice and Their Multilineage Differentiation Potentials

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: Long-Term Repopulation Patterns of CD34+/– MPL+/– SRCs in NOG Mice and Their Multilineage Differentiation Potentials

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Mouse Assay

    Primary and secondary human hematopoietic cell repopulation abilities of CD34 +/- MPL +/- SRCs. A total of 5× 10 3 18Lin-CD34 + MPL +/– cells (A) and 2 or 8×10 3 18Lin-CD34-MPL +/– cells (C) were transplanted into NOG mice. The percentages of human CD45 + cells in the right tibiae were serially analyzed at 5, 12, 18, and 22 to 24 weeks after transplantation. The data represent the mean±SD of the results from 9 to 13 mice at each time point. (B, D) The long-term (LT) human hematopoietic cell reconstitution in secondary NOG mice was also analyzed by the BM aspiration method at 12, 18, and 20 to 23 weeks after transplantation by six-color flow cytometry (FCM). The data represent the mean±SD of the results from 9 to 10 mice at each time point. * p

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: Primary and secondary human hematopoietic cell repopulation abilities of CD34 +/- MPL +/- SRCs. A total of 5× 10 3 18Lin-CD34 + MPL +/– cells (A) and 2 or 8×10 3 18Lin-CD34-MPL +/– cells (C) were transplanted into NOG mice. The percentages of human CD45 + cells in the right tibiae were serially analyzed at 5, 12, 18, and 22 to 24 weeks after transplantation. The data represent the mean±SD of the results from 9 to 13 mice at each time point. (B, D) The long-term (LT) human hematopoietic cell reconstitution in secondary NOG mice was also analyzed by the BM aspiration method at 12, 18, and 20 to 23 weeks after transplantation by six-color flow cytometry (FCM). The data represent the mean±SD of the results from 9 to 10 mice at each time point. * p

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Mouse Assay, Transplantation Assay, Flow Cytometry, Cytometry

    The frequency of severe combined immunodeficiency (SCID)-repopulating cells (SRCs) in 18Lin – CD34 +/– MPL +/– cells. Various numbers of 18Lin – CD34 + MPL + cells (50, 100, and 200, n = 18) (A), 18Lin – CD34 + MPL – cells (200, 400, and 800, n = 18) (B), 18Lin – CD34 – MPL + cells (400 and 2,000, n = 20) (C), and 18Lin – CD34 – MPL – cells (1,000, 1,600, and 3,000, n = 23) (D) were transplanted into NOG mice by intra-BM injection (IBMI). At week 12, the mice were euthanized, and the human CD45 + cell engraftment was analyzed. The solid line represents the estimated weighted mean frequency ( f WM ) of SRCs, and the upper and lower dotted lines represent the 95% confidence interval of f WM . The mice were scored as positive if more than 0.01% of the total murine BM cells were human CD45 + .

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: The frequency of severe combined immunodeficiency (SCID)-repopulating cells (SRCs) in 18Lin – CD34 +/– MPL +/– cells. Various numbers of 18Lin – CD34 + MPL + cells (50, 100, and 200, n = 18) (A), 18Lin – CD34 + MPL – cells (200, 400, and 800, n = 18) (B), 18Lin – CD34 – MPL + cells (400 and 2,000, n = 20) (C), and 18Lin – CD34 – MPL – cells (1,000, 1,600, and 3,000, n = 23) (D) were transplanted into NOG mice by intra-BM injection (IBMI). At week 12, the mice were euthanized, and the human CD45 + cell engraftment was analyzed. The solid line represents the estimated weighted mean frequency ( f WM ) of SRCs, and the upper and lower dotted lines represent the 95% confidence interval of f WM . The mice were scored as positive if more than 0.01% of the total murine BM cells were human CD45 + .

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Mouse Assay, Injection

    The isolation of 18Lin – CD34 +/– myeloproliferative leukemia virus (MPL) +/– cells from human cord blood (CB)-derived Lin – cells. Human CB-derived Lin – cells were stained with anti-18Lin, anti-CD34, anti-CD45, and anti-MPL monoclonal antibodies (mAbs). (A) The R1 gate was set on the blast–lymphocyte window. (B) The R2 gate was set on the 18Lin – living cells. (C) The cells in the R2 gate were subdivided into CD34 + (R3) and CD34 – (R4) fractions. (D, E) The CD34 +/– cells were further subdivided into MPL +/– (R5–R8) fractions. As shown in (D) and (E), the percentages of MPL + cells in the CD34 + (R5) and CD34 – (R7) fractions ranged from 13.6% to 56.4% (median: 36.9%, n = 9) and 1.4% to 33.1% (median: 10.3%, n = 9), respectively. 7AAD, 7-aminoactinomycin D; FSC, forward scatter; SSC, side scatter.

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: The isolation of 18Lin – CD34 +/– myeloproliferative leukemia virus (MPL) +/– cells from human cord blood (CB)-derived Lin – cells. Human CB-derived Lin – cells were stained with anti-18Lin, anti-CD34, anti-CD45, and anti-MPL monoclonal antibodies (mAbs). (A) The R1 gate was set on the blast–lymphocyte window. (B) The R2 gate was set on the 18Lin – living cells. (C) The cells in the R2 gate were subdivided into CD34 + (R3) and CD34 – (R4) fractions. (D, E) The CD34 +/– cells were further subdivided into MPL +/– (R5–R8) fractions. As shown in (D) and (E), the percentages of MPL + cells in the CD34 + (R5) and CD34 – (R7) fractions ranged from 13.6% to 56.4% (median: 36.9%, n = 9) and 1.4% to 33.1% (median: 10.3%, n = 9), respectively. 7AAD, 7-aminoactinomycin D; FSC, forward scatter; SSC, side scatter.

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Isolation, Derivative Assay, Staining

    The generation of CD34 + CD38 – CD45RA – CD90 + SRCs in a representative mouse that received human CB-derived 18Lin – CD34 – cells. Freshly sorted human CB-derived 18Lin – CD34 – cells were transplanted into NOG mice by IBMI. After 20 weeks, human CD45 + cell repopulation in the BM of a representative mouse was analyzed and is shown in the left column (A). Then 12Lin – /CD45RA – CD34 + CD38 – CD90 +/– cells were sorted (B–D). The 12 lineages contain CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD56, and CD235a. Next, these 12Lin – CD34 + CD38 – CD45RA – CD90 +/– cells (D) were transplanted into secondary NOG mice by IBMI, and the human cell repopulation was analyzed at 20 weeks after transplantation (E–G). Only the CD90 + cells repopulated in the secondary recipient NOG mice and showed a multilineage reconstitution. 7AAD, 7-Aminoactinomycin D.

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: The generation of CD34 + CD38 – CD45RA – CD90 + SRCs in a representative mouse that received human CB-derived 18Lin – CD34 – cells. Freshly sorted human CB-derived 18Lin – CD34 – cells were transplanted into NOG mice by IBMI. After 20 weeks, human CD45 + cell repopulation in the BM of a representative mouse was analyzed and is shown in the left column (A). Then 12Lin – /CD45RA – CD34 + CD38 – CD90 +/– cells were sorted (B–D). The 12 lineages contain CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD56, and CD235a. Next, these 12Lin – CD34 + CD38 – CD45RA – CD90 +/– cells (D) were transplanted into secondary NOG mice by IBMI, and the human cell repopulation was analyzed at 20 weeks after transplantation (E–G). Only the CD90 + cells repopulated in the secondary recipient NOG mice and showed a multilineage reconstitution. 7AAD, 7-Aminoactinomycin D.

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Derivative Assay, Mouse Assay, Transplantation Assay

    The colony-forming cell (CFC) capacities of 18Lin – CD34 +/– MPL +/– cells. (A) A total of 200 18Lin – CD34 +/– MPL +/– cells were cultured in the semisolid methylcellulose supplemented with 30% fetal calf serum (FCS) in the presence of six cytokines [thrombopoietin (THPO), stem cell factor (SCF), interleukin-3 (IL-3), granulocyte macrophage colony-stimulating factor (GM-CSF), G-CSF, and erythropoietin (Epo)] for 14 days or (B) supplemented with 10% platelet-poor plasma in the presence of three cytokines (THPO, IL-3, and Epo) for 10 days. The types of colonies were identified under inverted microscopy. The data represent the mean ± standard deviation (SD) of quadruple cultures. CFU-GM, colony forming unit-granulocyte/macrophage; BFU-E, erythroid burst-forming unit; CFU-Meg, megakaryocyte; CFU-Mix, erythrocyte-containing mixed; CFU-EM, erythrocyte/megakaryocyte mixed colony. * p

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: The colony-forming cell (CFC) capacities of 18Lin – CD34 +/– MPL +/– cells. (A) A total of 200 18Lin – CD34 +/– MPL +/– cells were cultured in the semisolid methylcellulose supplemented with 30% fetal calf serum (FCS) in the presence of six cytokines [thrombopoietin (THPO), stem cell factor (SCF), interleukin-3 (IL-3), granulocyte macrophage colony-stimulating factor (GM-CSF), G-CSF, and erythropoietin (Epo)] for 14 days or (B) supplemented with 10% platelet-poor plasma in the presence of three cytokines (THPO, IL-3, and Epo) for 10 days. The types of colonies were identified under inverted microscopy. The data represent the mean ± standard deviation (SD) of quadruple cultures. CFU-GM, colony forming unit-granulocyte/macrophage; BFU-E, erythroid burst-forming unit; CFU-Meg, megakaryocyte; CFU-Mix, erythrocyte-containing mixed; CFU-EM, erythrocyte/megakaryocyte mixed colony. * p

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Cell Culture, Inverted Microscopy, Standard Deviation

    Proposed model of the human CB-derived hematopoietic stem cell (HSC) hierarchy according to the CD34 and MPL expression pattern. According to our serial transplantation analyses, we propose that the expression pattern of CD34 antigen and MPL receptor defines/segregates human CB-derived primitive HSCs into CD34 – MPL + short-term (ST)-, CD34 + MPL + intermediate-term (IT)-, and CD34 +/– MPL – LT-HSCs, as shown in this figure. In our model, we use the following definition: ST-HSCs only support human cell repopulation in primary recipient mice; IT-HSCs support human cell repopulation in primary and secondary recipient mice; and LT-HSCs support human cell repopulation in primary, secondary, and tertiary recipient mice. The CD34 – MPL – LT-HSCs reside at the apex of the human HSC hierarchy. It is suggested that they first commit to CD34 + MPL – LT-HSCs, then commit to CD34 + MPL + IT-HSCs, and finally commit to CD34 – MPL + ST-HSCs.

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: Proposed model of the human CB-derived hematopoietic stem cell (HSC) hierarchy according to the CD34 and MPL expression pattern. According to our serial transplantation analyses, we propose that the expression pattern of CD34 antigen and MPL receptor defines/segregates human CB-derived primitive HSCs into CD34 – MPL + short-term (ST)-, CD34 + MPL + intermediate-term (IT)-, and CD34 +/– MPL – LT-HSCs, as shown in this figure. In our model, we use the following definition: ST-HSCs only support human cell repopulation in primary recipient mice; IT-HSCs support human cell repopulation in primary and secondary recipient mice; and LT-HSCs support human cell repopulation in primary, secondary, and tertiary recipient mice. The CD34 – MPL – LT-HSCs reside at the apex of the human HSC hierarchy. It is suggested that they first commit to CD34 + MPL – LT-HSCs, then commit to CD34 + MPL + IT-HSCs, and finally commit to CD34 – MPL + ST-HSCs.

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Derivative Assay, Expressing, Transplantation Assay, Mouse Assay

    The expression of Fms-like tyrosine kinase 3 (Flt3) on human CB-derived 18Lin – CD34 +/– MPL +/– cells. Human CB-derived Lin – cells were stained with anti-18Lin, anti-CD34, anti-CD45, anti-MPL, and anti-Flt3 mAbs. (A) The R1 gate was set on the blast– lymphocyte window. (B) The R2 gate was set on the 18Lin – living cells. (C) The cells in the R2 gate were subdivided into CD34 + (R3) and CD34 – (R4) fractions. (D, E) The CD34 +/– cells were further subdivided into four cell fractions according the expression of MPL and Flt3. The percentages of each fraction of cells are depicted in the figure.

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: The expression of Fms-like tyrosine kinase 3 (Flt3) on human CB-derived 18Lin – CD34 +/– MPL +/– cells. Human CB-derived Lin – cells were stained with anti-18Lin, anti-CD34, anti-CD45, anti-MPL, and anti-Flt3 mAbs. (A) The R1 gate was set on the blast– lymphocyte window. (B) The R2 gate was set on the 18Lin – living cells. (C) The cells in the R2 gate were subdivided into CD34 + (R3) and CD34 – (R4) fractions. (D, E) The CD34 +/– cells were further subdivided into four cell fractions according the expression of MPL and Flt3. The percentages of each fraction of cells are depicted in the figure.

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: Expressing, Derivative Assay, Staining

    In vitro lineage differentiation potentials of 18Lin – CD34 +/– MPL +/– cells and maintenance/generation of CD34 + cells from 18Lin – CD34 +/– MPL +/– cells in the coculture with bone marrow (BM)-derived mesenchymal stem cells (DP-MSCs). A total of 1,000 18Lin – CD34 +/– MPL +/– cells were cocultured with DP-MSCs for 7 days. (A) The fold increase in the total number of cells. (B) The absolute numbers of CD34 + cells maintained/generated in cocultures with DP-MSCs are shown. The absolute numbers of (C) CD33 + , (D) CD11b + , (E) CD14 + , and (F) CD41 + cells generated in cocultures are shown. The data represent the mean ± SD of quadruple cocultures. ** p

    Journal: Cell Transplantation

    Article Title: CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

    doi: 10.3727/096368916X694201

    Figure Lengend Snippet: In vitro lineage differentiation potentials of 18Lin – CD34 +/– MPL +/– cells and maintenance/generation of CD34 + cells from 18Lin – CD34 +/– MPL +/– cells in the coculture with bone marrow (BM)-derived mesenchymal stem cells (DP-MSCs). A total of 1,000 18Lin – CD34 +/– MPL +/– cells were cocultured with DP-MSCs for 7 days. (A) The fold increase in the total number of cells. (B) The absolute numbers of CD34 + cells maintained/generated in cocultures with DP-MSCs are shown. The absolute numbers of (C) CD33 + , (D) CD11b + , (E) CD14 + , and (F) CD41 + cells generated in cocultures are shown. The data represent the mean ± SD of quadruple cocultures. ** p

    Article Snippet: The collected BM cells were stained with a cocktail of mAbs [mouse CD45.1 (Beckman Coulter), CD19, CD33 (Beckman Coulter), and CD34 (BD Bioscience) and human CD45 (BioLegend)], and the human cell repopulation ability was analyzed by FCM (FACSCantoII; BD Bioscience).

    Techniques: In Vitro, Derivative Assay, Generated

    Characterization of rat bone marrow-derived mesenchymal stromal cells (BMSCs). (a) The plastic-adherent cells demonstrated a homogeneous fibroblast-like and spindle-shaped morphology. Original magnification, ×100. (b) Adipogenic differentiation of BMSCs stained with oil red O. Original magnification, ×200. (c) Osteogenic differentiation of BM-MSCs stained with alizarin red. Original magnification, ×400. (d) FACS analysis demonstrated expression of markers attributed to BMSCs. The cells were devoid of hematopoietic cells as indicated by the lack of CD45 and CD34. The MSC-specific markers, CD29, CD44, CD73, CD105, and CD90 were strongly expressed on the cells.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Thioredoxin-1 Protects Bone Marrow-Derived Mesenchymal Stromal Cells from Hyperoxia-Induced Injury In Vitro

    doi: 10.1155/2018/1023025

    Figure Lengend Snippet: Characterization of rat bone marrow-derived mesenchymal stromal cells (BMSCs). (a) The plastic-adherent cells demonstrated a homogeneous fibroblast-like and spindle-shaped morphology. Original magnification, ×100. (b) Adipogenic differentiation of BMSCs stained with oil red O. Original magnification, ×200. (c) Osteogenic differentiation of BM-MSCs stained with alizarin red. Original magnification, ×400. (d) FACS analysis demonstrated expression of markers attributed to BMSCs. The cells were devoid of hematopoietic cells as indicated by the lack of CD45 and CD34. The MSC-specific markers, CD29, CD44, CD73, CD105, and CD90 were strongly expressed on the cells.

    Article Snippet: Phycoerythrin- (PE-) coupled antibodies against CD29 (eBiosciences, cat. no. 12-0291, San Diego, CA, USA), CD34 (Santa Cruz Biotechnology, cat. no. sc-74499), CD44 (Santa Cruz Biotechnology, cat. no. sc-7297), CD45 (Santa Cruz Biotechnology, cat. no. sc-1178), and CD90 (Santa Cruz Biotechnology, cat. no. sc-53456) were added separately, followed by incubation at 4°C for 30 minutes.

    Techniques: Derivative Assay, Staining, FACS, Expressing

    Hypoxia reduces proliferation of CD34 + hematopoietic stem and progenitor cells. CD34 + and CD133 + cells in cultures in both hypoxia and normoxia were determined by flow cytometry on days 1, 7, 14 and 21. (A) Fold change in the number of CD34 + cells in the normoxic and hypoxic cultures was calculated relative to day 1. (B) Fold change in the number of CD133 + cells in the normoxic and hypoxic cultures was calculated relative to day 1. (C) Percentage of CD34 + cells in the Lin − /Live population is shown. Data are represented as the mean with standard error ( n = 6 for CD34 and n = 4 for CD133). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Hypoxia reduces proliferation of CD34 + hematopoietic stem and progenitor cells. CD34 + and CD133 + cells in cultures in both hypoxia and normoxia were determined by flow cytometry on days 1, 7, 14 and 21. (A) Fold change in the number of CD34 + cells in the normoxic and hypoxic cultures was calculated relative to day 1. (B) Fold change in the number of CD133 + cells in the normoxic and hypoxic cultures was calculated relative to day 1. (C) Percentage of CD34 + cells in the Lin − /Live population is shown. Data are represented as the mean with standard error ( n = 6 for CD34 and n = 4 for CD133). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Flow Cytometry, MANN-WHITNEY

    Hypoxia increases erythroid differentiation. (A) Immunophenotypic analysis of lineage cells: monocytes (CD34 − /CD14 + /CD66b − ), granulocytes (CD34 − /CD14 − /CD66b + ), megakaryocytes (CD34 − /CD41a + /CD235a − ), and erythrocytes (CD34 − /CD41a − /CD235a + ) on days 14 and 21. Percentage of positive cells in the CD34 − /Live population are illustrated. (B) Distribution of cells expressing CD33 was determined on days 1, 7, 14, and 21 by flow cytometry. Percentages of total CD33 + , CD34 + /CD33 + , and CD34 − /CD33 + cells in the live population are illustrated. (C) Distribution of cells expressing CD11b was determined on days 1, 7, 14, and 21 by flow cytometry. Percentages of total CD11b + , CD34 + /CD11b + , and CD34 − /CD11b + cells in the live population are illustrated. Data are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Hypoxia increases erythroid differentiation. (A) Immunophenotypic analysis of lineage cells: monocytes (CD34 − /CD14 + /CD66b − ), granulocytes (CD34 − /CD14 − /CD66b + ), megakaryocytes (CD34 − /CD41a + /CD235a − ), and erythrocytes (CD34 − /CD41a − /CD235a + ) on days 14 and 21. Percentage of positive cells in the CD34 − /Live population are illustrated. (B) Distribution of cells expressing CD33 was determined on days 1, 7, 14, and 21 by flow cytometry. Percentages of total CD33 + , CD34 + /CD33 + , and CD34 − /CD33 + cells in the live population are illustrated. (C) Distribution of cells expressing CD11b was determined on days 1, 7, 14, and 21 by flow cytometry. Percentages of total CD11b + , CD34 + /CD11b + , and CD34 − /CD11b + cells in the live population are illustrated. Data are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Expressing, Flow Cytometry, MANN-WHITNEY

    Hypoxia enhances expression of erythroid markers. (A) Longitudinal analysis of CD71 relative to the erythroid marker CD235a in cultures incubated in normoxia or hypoxia. Percentages of CD71 + /CD235a − , CD71 − /CD235a + , and CD71 + /CD235a + in the CD34 − /Live population are illustrated. (B) Longitudinal analysis of CD71 relative to the erythroid marker CD239 in cultures incubated in normoxia or hypoxia. Percentages of CD71 + /CD239 − , CD71 − /CD239 + , and CD71 + /CD239 + cells in the CD34 − /Live population are illustrated. Data are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. (C) tSNE plots revealing distribution of CD71 + , CD235a + , and CD239 + cells on day 21 in normoxia or hypoxia. (D) tSNE plots of overlay of CD71 + , CD235a + , and CD239 + cells in normoxia or hypoxia on day 21. tSNE analysis was performed in FlowJo. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Hypoxia enhances expression of erythroid markers. (A) Longitudinal analysis of CD71 relative to the erythroid marker CD235a in cultures incubated in normoxia or hypoxia. Percentages of CD71 + /CD235a − , CD71 − /CD235a + , and CD71 + /CD235a + in the CD34 − /Live population are illustrated. (B) Longitudinal analysis of CD71 relative to the erythroid marker CD239 in cultures incubated in normoxia or hypoxia. Percentages of CD71 + /CD239 − , CD71 − /CD239 + , and CD71 + /CD239 + cells in the CD34 − /Live population are illustrated. Data are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. (C) tSNE plots revealing distribution of CD71 + , CD235a + , and CD239 + cells on day 21 in normoxia or hypoxia. (D) tSNE plots of overlay of CD71 + , CD235a + , and CD239 + cells in normoxia or hypoxia on day 21. tSNE analysis was performed in FlowJo. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Expressing, Marker, Incubation, MANN-WHITNEY

    Hypoxia promotes the development of megakaryoerythroid progenitors. (A) Schematic of myeloerythroid differentiation from hematopoietic stem cells. The surface markers used for immunophenotyping of the cells interrogated in this study are indicated. (B) Percentage of multipotent progenitors (MPP), common myeloid progenitors (CMP), megakaryoerythroid progenitors (MEP), and granulocyte−monocyte progenitors (GMP) in cultures incubated in hypoxia and normoxia. For MPPs, the percentage of positive cells in the Lin − /Live population was determined, and for CMPs, GMPs, and MEPs, the percentage of positive cells in the MPP population was determined on days 1, 7, 14, and 21. Data are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. (C) tSNE analysis was performed on flow cytometry standard files. tSNE plots for day 21 analysis revealing CD34 + and CD38 + cells in normoxia and hypoxia are represented in the Lin − /Live population. (D) tSNE analysis plots revealing the distribution of CMPs, GMPs, and MEPs in the CD34 + /CD38 + population in normoxia or hypoxia on day 21 are represented. (E) Histograms for CD45Ra and CD123 expression indicating relative distribution of CMPs (CD45Ra − /CD123 lo ), GMPs (CD45Ra + /CD123 lo ), and MEPs (CD45Ra − /CD123 − ) in normoxia and hypoxia. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Hypoxia promotes the development of megakaryoerythroid progenitors. (A) Schematic of myeloerythroid differentiation from hematopoietic stem cells. The surface markers used for immunophenotyping of the cells interrogated in this study are indicated. (B) Percentage of multipotent progenitors (MPP), common myeloid progenitors (CMP), megakaryoerythroid progenitors (MEP), and granulocyte−monocyte progenitors (GMP) in cultures incubated in hypoxia and normoxia. For MPPs, the percentage of positive cells in the Lin − /Live population was determined, and for CMPs, GMPs, and MEPs, the percentage of positive cells in the MPP population was determined on days 1, 7, 14, and 21. Data are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. (C) tSNE analysis was performed on flow cytometry standard files. tSNE plots for day 21 analysis revealing CD34 + and CD38 + cells in normoxia and hypoxia are represented in the Lin − /Live population. (D) tSNE analysis plots revealing the distribution of CMPs, GMPs, and MEPs in the CD34 + /CD38 + population in normoxia or hypoxia on day 21 are represented. (E) Histograms for CD45Ra and CD123 expression indicating relative distribution of CMPs (CD45Ra − /CD123 lo ), GMPs (CD45Ra + /CD123 lo ), and MEPs (CD45Ra − /CD123 − ) in normoxia and hypoxia. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Incubation, MANN-WHITNEY, Flow Cytometry, Expressing

    Hypoxia causes a metabolic shift in CD34 + hematopoietic stem and progenitor cells. (A) Principal component analysis of samples on the basis of fold change in metabolites in spent medium relative to fresh medium from CD34 + cell cultures incubated in hypoxia or normoxia. (B) Fold change in glucose and lactate in spent medium relative to fresh medium from CD34 + cell cultures incubated in normoxia or hypoxia. (C) Fold change in amino acids in spent medium relative to fresh medium from CD34 + cell cultures incubated in normoxia or hypoxia. Spent medium was collected on day 4 of culture. Statistical analysis was performed in MetaboAnalyst, and p values ≤ 0.05 were considered significant. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Hypoxia causes a metabolic shift in CD34 + hematopoietic stem and progenitor cells. (A) Principal component analysis of samples on the basis of fold change in metabolites in spent medium relative to fresh medium from CD34 + cell cultures incubated in hypoxia or normoxia. (B) Fold change in glucose and lactate in spent medium relative to fresh medium from CD34 + cell cultures incubated in normoxia or hypoxia. (C) Fold change in amino acids in spent medium relative to fresh medium from CD34 + cell cultures incubated in normoxia or hypoxia. Spent medium was collected on day 4 of culture. Statistical analysis was performed in MetaboAnalyst, and p values ≤ 0.05 were considered significant. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Incubation

    Hypoxia promotes enucleation of erythroid cells. (A) Immunoblotting of α - and β -hemoglobin expression in CD34 + cell cultures incubated in normoxia or hypoxia and in cord blood-derived mononuclear cells (MNCs). Average protein expression (§SE, n = 3) normalized to GAPDH is illustrated. (B) Enucleation was determined by SYTO16 staining. Contour plots for CD235a + and SYTO16 + cells in cultures incubated in hypoxia or normoxia. (C) Percentages of enucleated (CD235a + /SYTO16 − ) and nucleated (CD235a + /SYTO16 + ) erythroid cells in the CD34 − /Live population on days 14 and 21 are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Hypoxia promotes enucleation of erythroid cells. (A) Immunoblotting of α - and β -hemoglobin expression in CD34 + cell cultures incubated in normoxia or hypoxia and in cord blood-derived mononuclear cells (MNCs). Average protein expression (§SE, n = 3) normalized to GAPDH is illustrated. (B) Enucleation was determined by SYTO16 staining. Contour plots for CD235a + and SYTO16 + cells in cultures incubated in hypoxia or normoxia. (C) Percentages of enucleated (CD235a + /SYTO16 − ) and nucleated (CD235a + /SYTO16 + ) erythroid cells in the CD34 − /Live population on days 14 and 21 are represented as the mean with standard error ( n = 4). Statistical analysis was performed using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Expressing, Incubation, Derivative Assay, Staining, MANN-WHITNEY

    Expression of CD105 is persistent in hypoxia. (A) Longitudinal analysis of CD71 and CD105 in cultures incubated in normoxia or hypoxia. Percentages of CD71 + /CD105 − , CD71 − /CD105 + , and CD71 + /CD105 + in the CD34 − /Live population are illustrated. (B) Longitudinal analysis of CD105 relative to the erythroid marker CD235a in cultures incubated in normoxia or hypoxia. Percentages of CD105 + /CD235a − , CD105 − /CD235a + , and CD105 + /CD235s + cells in the CD34 − /Live population are illustrated. Data are represented as the mean with standard error ( n = 4). Statistical significance was calculated using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. (C) tSNE plots revealing the distribution of CD105 + cells and overlay of CD71 + , CD105 + , and CD235a + cells on day 21 in normoxia or hypoxia. tSNE analysis was performed in FlowJo. (D) Longitudinal analysis of the CD49d and CD233 in cultures incubated in normoxia or hypoxia. Percentages of CD49d − /CD233 − , CD49d + /CD233 − , CD49d + /CD233 + , and CD49d + /CD233 − cells in the CD235a + population are illustrated. * p

    Journal: Experimental hematology

    Article Title: Hypoxia promotes erythroid differentiation through the development of progenitors and proerythroblasts

    doi: 10.1016/j.exphem.2021.02.012

    Figure Lengend Snippet: Expression of CD105 is persistent in hypoxia. (A) Longitudinal analysis of CD71 and CD105 in cultures incubated in normoxia or hypoxia. Percentages of CD71 + /CD105 − , CD71 − /CD105 + , and CD71 + /CD105 + in the CD34 − /Live population are illustrated. (B) Longitudinal analysis of CD105 relative to the erythroid marker CD235a in cultures incubated in normoxia or hypoxia. Percentages of CD105 + /CD235a − , CD105 − /CD235a + , and CD105 + /CD235s + cells in the CD34 − /Live population are illustrated. Data are represented as the mean with standard error ( n = 4). Statistical significance was calculated using the Mann-Whitney test, and p values ≤ 0.05 were considered significant. (C) tSNE plots revealing the distribution of CD105 + cells and overlay of CD71 + , CD105 + , and CD235a + cells on day 21 in normoxia or hypoxia. tSNE analysis was performed in FlowJo. (D) Longitudinal analysis of the CD49d and CD233 in cultures incubated in normoxia or hypoxia. Percentages of CD49d − /CD233 − , CD49d + /CD233 − , CD49d + /CD233 + , and CD49d + /CD233 − cells in the CD235a + population are illustrated. * p

    Article Snippet: For enucleation analysis, cells were stained with antibodies to CD34 (APC-Cy7) and CD235a (APC) and washed as described above, and then incubated with 100 nmol/L SYTO16 (excitation/emission 488/518 nm, Thermo Fisher Scientific), a live cell permeant dye, for 15 min at room temperature [ ].

    Techniques: Expressing, Incubation, Marker, MANN-WHITNEY