Journal: Genes & Cancer

Article Title: CD133-positive cancer stem cells from Colo205 human colon adenocarcinoma cell line show resistance to chemotherapy and display a specific metabolomic profile

doi:

Figure Lengend Snippet: A-B. Cytometry analyses of Nestin, CK20, CD133, Oct3/4, CD44 and CD44v9 on 10% FBS Colo205 growing cells (control) and serum-free growing cultures (week 1 to week 5).

Article Snippet: The purity of sorted cells was evaluated by flow cytometry using FACSCalibur (BD Biosciences) after labeling with anti-human CD133/2 or CD44 antibody (Miltenyi Biotec).

Techniques: Cytometry, Control

Journal: Genes & Cancer

Article Title: CD133-positive cancer stem cells from Colo205 human colon adenocarcinoma cell line show resistance to chemotherapy and display a specific metabolomic profile

doi:

Figure Lengend Snippet: A. Tumorsphere evaluation of CD133+ and CD133- sorted cells. B. Invasiveness of different cell fractions measured using a collagen-based invasion kit (relative DO measured at 560 nm). C. Survival assay after chemotherapy treatment with cisplatin and 5-FU. D. Metabolite quantification after CE-TOF-MS experiments. Only metabolites of interest for Colo205 cells cultured in 10% FBS (control; green bar), CD133+ sorted cells (red bar) and CD44+ sorted cells (yellow bar) are reported here.

Article Snippet: The purity of sorted cells was evaluated by flow cytometry using FACSCalibur (BD Biosciences) after labeling with anti-human CD133/2 or CD44 antibody (Miltenyi Biotec).

Techniques: Clonogenic Cell Survival Assay, Cell Culture, Control

Journal: Phytomedicine

Article Title: Exploring the Therapeutic Potential of HAPC in COVID-19-Induced Acute Lung Injury

doi: 10.1016/j.phymed.2025.156563

Figure Lengend Snippet: Fig. 3. Optimization of chicoric acid to target CD44 using bioengineering nanospores. (A,B) Demonstration of an easy synthesis strategy for HAPC nano particles using an emulsion method. (C, D) Lactobacillus murinus conidia cell wall (CW). (E) Validation of the assembly of CW and HAPC nanoparticles via FRET. (F) Size distribution of HAPC nanoparticles. (G) Size distribution of optimized HAPC nanoparticles. H) Interaction between CW and PEI identified via FRET.

Article Snippet: HAPC–Cy5.5, 50-μg HAPC–Cy5.5 plus 100-μg HA, or 50-μg seng-cy5.5 and 5-μl anti-CD44 antibody in a serum-free medium for 2 h RAW 264.7 cells (1 × 105 cells/well) were treated with 100-ng/ml LPS and 10-ng/ml IFN-γ (HY-P7025, MCE) for 24 h to activate macrophages.

Techniques: Emulsion, Biomarker Discovery

Journal: International journal of molecular medicine

Article Title: Autophagy is essential for the endothelial differentiation of breast cancer stem‑like cells.

doi: 10.3892/ijmm.2019.4399

Figure Lengend Snippet: Figure 1. Flow cytometry and microsphere culture of BCSLCs. (A) CD24‑/low cells were isolated from MCF‑7 using immunomagnetic beads, and the expres- sion of CD24 in MCF‑7 and isolated CD24‑/low cells were assessed using flow cytometry. (a) Isotype control for (b); (b) MCF‑7 cells; (c) isotype control for (d); (d) isolated CD24‑/low cells. (B) CD44+ cells were further isolated from CD24‑/low cells and the expression of CD44 was assessed using flow cytometry. (a) Isotype control for (b); (b) CD44+ cells. (C) Expression of CD24 and CD44 in MCF‑7, BCSLCs and BCSLCs after eight passages. (a) isotype control for (b); (b) MCF‑7 cells; (c) CSLCs; (d) isotype control for (e); (e) BCSLCs after eight passages. (D) The isolated BCCSLCs were cultured in microspheres for 0 and 64 h in stem cell culture medium. BCSLC, breast cancer stem‑like cell; PE, phycoerythrin.

Article Snippet: The following primary antibodies were used: Fluorescein isothiocyanate (FITc)-conjugated anti-human cd44 (cat. no. 130-113-903; Miltenyi Biotec GmbH), phycoerythrin (PE)-conjugated anti-human cd24 (cat. no. 130-098-861; Miltenyi Biotec GmbH) PE-conjugated anti-human cd31 (cat. no. 130-110-807; Miltenyi Biotec GmbH) and FITc-conjugated anti-human cd105 (cat. no. 130-098-778; Miltenyi Biotec GmbH).

Techniques: Flow Cytometry, Isolation, Control, Expressing, Cell Culture, Stem Cell Culture

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: (A) Free-floating tumor spheres were formed from EBV-positive C666-1 cells (left panel) and they demonstrated ability to differentiate comparably to monolayer cells upon administering complete medium (right panel). (B) By qRT-PCR, multiple stem cell-related genes (OCT4, NANOG, ALDH1, CKIT, CD44, CD133) were enriched in spheroids when compared to parental C666-1. Transcription of SOX2 was not increased in the spheroids. (C) SOX2 protein was frequently expressed in C666-1 sphere-forming cells. By flow cytometry, SOX2-positive (SOX2+) cells were found to be enriched in and constituted over 60% of sphere-forming cell population. (D) Over 80% of sphere-forming cells expressed cell surface marker CD44 while CD44+ cells in detected in parental C666-1 and other NPC xenografts were significantly lower (all P <0.001). Histograms denoting mean ± SE (n≥3) with statistical significance calculated by t-test (* P <0.05, ** P <0.01, *** P <0.001).

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Quantitative RT-PCR, Flow Cytometry, Marker

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: In vivo tumorigenic capacity of sphere-forming cells and unselected parental cells of C666-1 in nude mice.

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: In Vivo

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: By flow cytometry, SOX2 was found to be preferentially expressed on CD44+ cells and coincidentally, SOX2 expression was rarely detected in CD44− cells. Cells coexpressing both CD44 and SOX2 were found to be enriched in spheroids. Histograms denoting mean ± SE (n≥3) with statistical significance calculated by t-test (* P <0.05, ** P <0.01, *** P <0.001).

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Flow Cytometry, Expressing

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: CD44+ cell fraction exhibited a significantly higher (A) clone formation efficiency and (B) sphere-forming efficiency when compared to CD44− cell fraction. In addition, (C) CD44+ cells exhibited significantly higher proliferation rate than CD44− cells. (D) Developmental hierarchy feature of CD44+ cells. Percentage of CD44+ cells were continually reduced in the isolated CD44+ cell fraction over time. (E) CD44+ cells exhibited higher resistance to 5-FU treatment when compared to the CD44− and parental C666-1 cells. All graphs denoting mean ± SE (n≥3) with statistical significance calculated by t-test (* P <0.05, ** P <0.01, *** P <0.001).

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Isolation

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: (A) By qRT-PCR, multiple EBV genes (EBER, BARF1, LMP1, LMP2A, EBNA1 and BZLF1) were found to be overexpressed in spheroids when compared to monolayer C666-1 cells. EBV copy number in these cells was determined by qPCR. (B) Selected genes aberrantly expressed in spheroids were confirmed by qRT-PCR. The significantly upregulated genes include chemokines and receptors (CCR7, CCL4, CX3CL1 and IL-8), cell adhesion molecule SELE, signaling molecules (GLI1, FOXN4) and ABC transporters (ABCC3, ABCC11). (C) Cell surface-expressed CCR7 was found to be frequently expressed in sphere-forming cells (>60%) by flow cytometry. The CCR7+ cell subpopulation was also detected in NPC lines and primary tumors (<5%). (D) CD44+CCR7+cells were also found to be enriched in spheroids. Histograms denoting mean ± SE (n≥3) with statistical significance calculated by t-test (* P <0.05, ** P <0.01, *** P <0.001).

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Quantitative RT-PCR, Flow Cytometry

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: Selection of aberrantly expressed genes in sphere-forming cells compared to monolayer C666-1 cells.

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Selection, Significance Assay, Variant Assay, Binding Assay, Histone Deacetylase Assay, RNA Binding Assay

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: Representative primary NPC cases with high (A), medium (B), low (C) expression of CCR7. (D) Primary NPC with absence of CCR7 expression was shown. CCR7 staining were detected in few infiltrating lymphocytes, but not in the tumor cells. Primary tumors with high (E) and medium (F) CD44 expression were shown. In (G) and (H), weak CD44 expression was detected in the tumor cells while strong CD44 staining in infiltrating lymphocytes was commonly found.

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Expressing, Staining

Journal: PLoS ONE

Article Title: CD44+ Cancer Stem-Like Cells in EBV-Associated Nasopharyngeal Carcinoma

doi: 10.1371/journal.pone.0052426

Figure Lengend Snippet: To evaluate the function of CCR7 in CSCs, C666-1 was treated with CCR7 blocking antibody and its proliferation, clone-forming and sphere-forming efficiency were investigated. (A) Proliferation of CD44+ cells was inhibited after treatment with CCR7 blocking antibody. (B) The clone formation efficiency of C666-1 cells was diminished after CCR7 blocking and (C) the spheroid-forming ability was significantly inihibited ( P <0.001) when compared to untreated controls. Histograms denoting mean ± SE (n≥3) with statistical significance calculated by t-test (* P <0.05, *** P <0.001).

Article Snippet: CD44-positive (CD44+) and CD44-negative (CD44−) C666-1 cells were separated by using anti-CD44 magnetic bead-coupled antibody and the magnetic-activated cell sorting (MACS) system (Miltenyi Biotec).

Techniques: Blocking Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting stem-property and vasculogenic mimicry for sensitizing paclitaxel therapy of triple-negative breast cancer by biomimetic codelivery

doi: 10.1016/j.apsb.2025.04.006

Figure Lengend Snippet: Regulation of the stem-property in 4T1 cells. (A) Transwell analysis in 4T1 cells, scale bars = 200 μm; (B, C) The representative images of scratch assay in 4T1 cells and the statistical analysis of wound area, scale bar = 100 μm; (D) Representative images of the 4T1 tumorspheres on Day 12 after different treatments, scale bar = 50 μm; (E, F) The statistical analysis of number and diameter of tumorspheres; (G) Schematic mechanism of SHK inhibiting stem-property of 4T1 cells; (H) Western blot analysis of PKM2, β -Catenin, epithelial or mesenchymal markers, stem-property markers in 4T1 cells; Quantization of Western blot analysis (I) and the mRNA levels (J) of β- catenin, Pkm2, Vimentin, Oct4, Sox2, Nanog and E-cadherin in 4T1 cells; (K, L) The population of CD44 + /CD24 ‒ subtype in drug treated 4T1 cells and statistical analysis. Data are presented as mean ± SD ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns, not significant.

Article Snippet: MHC II antibody, CD206 antibody, CD44 antibody, CD24 antibody were purchased from Elabscience (Wuhan, China).

Techniques: Wound Healing Assay, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting stem-property and vasculogenic mimicry for sensitizing paclitaxel therapy of triple-negative breast cancer by biomimetic codelivery

doi: 10.1016/j.apsb.2025.04.006

Figure Lengend Snippet: In vivo therapeutic efficacy of nanoparticles in a mouse model with orthotopic breast cancer. (A) Therapeutic schedule; (B) The tumor image, (C) tumor volume curves and (D) tumor weight, (E) Body weight curves; (F) Statistical analysis of body weight on the last day during treatment; (G) The ratio of the M1-like TAM (F4/80 + CD86 + ) subset; (H) The ratio of the M2-like TAM (F4/80 + CD206 + ) subset; (I) The ratio of the matured DC (CD80 + CD86 + ) subset; (J) The ratio of the PMN-MDSC (Ly6G + Ly6C − ) subset; (K) The ratio of the memory T cells (CD62L + CD44 + ) subset; (L) The ratio of the CD8 + T cells (CD3 + CD8 + ) subset; (M, N) The ratio of the cytotoxicity CD8 + T cells (IFN γ + /Granzyme B + ) subset. Data are presented as mean ± SD ( n = 3–6). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns, not significant.

Article Snippet: MHC II antibody, CD206 antibody, CD44 antibody, CD24 antibody were purchased from Elabscience (Wuhan, China).

Techniques: In Vivo, Drug discovery