mouse bone marrow mesenchymal stem cells bmscs  (ATCC)

Journal: Bioactive Materials

Article Title: An engineered M2 macrophage-derived exosomes-loaded electrospun biomimetic periosteum promotes cell recruitment, immunoregulation, and angiogenesis in bone regeneration

doi: 10.1016/j.bioactmat.2025.03.027

Figure Lengend Snippet: Construction of Engineered Exos. (A) Representative confocal microscopic images of BMSC Apts after co-culture with BMSCs for 4 h. Red: AF647-labeled Apt; green: F-actin; blue: DAPI-labeled cell nuclei. (B) Flow cytometry analysis was performed to detect the specificity of AF647-labeled BMSC Apts. (C) Flow cytometry analysis of M2 macrophage markers F4/80 and CD206. (D) Confocal microscopic image of DiO-labeled Exos cocultured with BMSCs. (E) WB analysis of exosomal positive and negative markers in Exos and donor cells. (F) Nanoflow cytometry analysis of the binding of Apt nanoparticles to Exos. (G) WB analysis of exosomal positive and negative markers in Exo and Apt-NP-Exo groups. (H) Nanoflow cytometry analysis of size distribution of Exos and Apt-NP-Exos. (I) TEM images of Exos and Apt-NP-Exos.

Article Snippet: To verify the specificity of the Apt for BMSCs, mouse BMSCs (Procell, China) were seeded in 15 mm confocal culture dishes and cultured in Dulbecco's modified Eagle medium (DMEM, Gibco, USA) supplemented with 10 % fetal bovine serum (FBS, ExCell, China) and 1 % penicillin-streptomycin (PS, Gibco, USA) at 37 °C under 5 % CO 2 .

Techniques: Co-Culture Assay, Labeling, Flow Cytometry, Cytometry, Binding Assay

Journal: Bioactive Materials

Article Title: An engineered M2 macrophage-derived exosomes-loaded electrospun biomimetic periosteum promotes cell recruitment, immunoregulation, and angiogenesis in bone regeneration

doi: 10.1016/j.bioactmat.2025.03.027

Figure Lengend Snippet: Biological Effects of Engineered Exos on BMSCs. (A) Representative confocal microscopic images and (B) quantitative analysis showed the internalization of Exos, NP-Exos, and Apt-NP-Exos after culture with BMSCs for 4 h. Red: AF647-labeled NP or Apt-NP; green: DiO-labeled Exos; blue: DAPI-labeled cell nuclei. (C) CCK-8 assay was conducted to evaluate the proliferation of BMSCs treated with different concentrations of Apt-NP-Exo at 24 and 48 h. (D) ALP staining of BMSCs on day 7 and ARS staining on day 14. (E) Quantification of ARS staining. (F) ALP activity of BMSCs on day 7. (G) WB assay and (H) quantitative analysis of Runx2 and BMP-2 expression in BMSCs. Data were shown as mean ± SDs. ns, no significance, ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: To verify the specificity of the Apt for BMSCs, mouse BMSCs (Procell, China) were seeded in 15 mm confocal culture dishes and cultured in Dulbecco's modified Eagle medium (DMEM, Gibco, USA) supplemented with 10 % fetal bovine serum (FBS, ExCell, China) and 1 % penicillin-streptomycin (PS, Gibco, USA) at 37 °C under 5 % CO 2 .

Techniques: Labeling, CCK-8 Assay, Staining, Activity Assay, Expressing

Journal: Bioactive Materials

Article Title: An engineered M2 macrophage-derived exosomes-loaded electrospun biomimetic periosteum promotes cell recruitment, immunoregulation, and angiogenesis in bone regeneration

doi: 10.1016/j.bioactmat.2025.03.027

Figure Lengend Snippet: Preparation of PEC-Apt-NP-Exo and Its Effect on Osteogenic Differentiation of BMSCs. (A) Schematic diagram illustrating the preparation of PEC. (B) Comparative study of femoral periosteum and periosteal ECM through HE, Masson, and DAPI staining analysis. (C) Agarose gel electrophoresis and (D) quantification of DNA content in femoral periosteum and periosteal ECM. (E) Confocal microscopy images and (F) quantification were utilized to evaluate the effectiveness of EDC/NHS in coating engineered Exos onto PECs via catalyzed amide condensation reactions. (G) Confocal microscopy images of core-shell structures. (H) Fluorescence microplate reader assay to evaluate the sustained release of engineered Exos. (I) CCK-8 assay and (J) live-dead staining were conducted to evaluate the effect of various membranes on BMSC growth. Green: calcein-AM, living cells; red: PI, dead cells (K) ALP staining of BMSCs on day 7 and ARS staining on day 14. (L) ALP activity of BMSCs on day 7. (M) Quantification of ARS staining. (N) WB assay and (O, P) quantitative analysis of Runx2 and OPN expression in BMSCs. Data were shown as mean ± SDs. ns, no significance, ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: To verify the specificity of the Apt for BMSCs, mouse BMSCs (Procell, China) were seeded in 15 mm confocal culture dishes and cultured in Dulbecco's modified Eagle medium (DMEM, Gibco, USA) supplemented with 10 % fetal bovine serum (FBS, ExCell, China) and 1 % penicillin-streptomycin (PS, Gibco, USA) at 37 °C under 5 % CO 2 .

Techniques: Staining, Agarose Gel Electrophoresis, Confocal Microscopy, Fluorescence, CCK-8 Assay, Activity Assay, Expressing