Journal: Advanced Healthcare Materials

Article Title: Fibrillar Bundles as Fibrous Filler Materials for Attaining Cell Anisotropy in Bioprinting

doi: 10.1002/adhm.202503767

Figure Lengend Snippet: Bioprinting composite bioinks containing encapsulated C2C12 myoblast cells in GelMA with 3.75% concentration. a) Schematic representation of GelMA composite bioink preparation. b) Quantification of the cell viability after 3D bioprinting of the composite bioink using live/dead assay and fluorescent microscopy imaging ( calcein AM (green) and ethidium homodimer‐1 (red)) with and without the fibrillar bundles. Fiber caps are spotted with red color due to the autofluorescence. Monitoring the cell behavior using green cell tracker‐stained myoblasts and fluorescence microscopy images (n=3), c) directly, and d) three days after 3D bioprinting. e) a larger snapshot of the fully cell‐covered bundle fiber cultured for three days after 3D bioprinting.

Article Snippet: Instead, Cell‐tracker green CMFDA was purchased from Invitrogen, Insulin‐transferrin‐selenium Solution (ITS), Fetal calf serum (FCS), horse serum (HS), penicillin−streptomycin (pen/strep), and GlutaMax were purchased from Gibco.

Techniques: Concentration Assay, Live Dead Assay, Microscopy, Imaging, Staining, Fluorescence, Cell Culture

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Sequential MR Image-Guided Local Immune Checkpoint Blockade Cancer Immunotherapy Using Ferumoxytol Capped Ultralarge Pore Mesoporous Silica Carriers after Standard Chemotherapy

doi: 10.1002/smll.201904378

Figure Lengend Snippet: Immunogenic cell death induction (ICD) of Tramp C1 and followed adaptive immune responses with PD-L1 overexpression. A) Schematic illustration of ICD and following CRT, HMGB1, and PD-L1 expression. B,C) Validation of ICD after Cbz treatment. 1 × 106 Tramp C1 cells were cultured with 0.5 × 10−6 m of Cbz for 18 h. CRT and HMGB1, immunogenic cell death markers were analyzed by ELISA and flow cytometry. D) In vivo DC maturation test after Cbz treatment. Tramp C1 prostate tumor bearing mice were treated with PBS as negative control and Cbz only for ICD induction. Splenocytes were isolated and prepared with CD11c as DC marker, CD80, and CD86 as DC maturation marker. Data were obtained and analyzed from CD11c+ gate by flow cytometry. E) Production of TNF-α, IL-12p70 and IL-10 from ICD induced DCs after ex vivo restimulation. Cytokine profiles were determined by ELISA. F) Tumor specific T cell activation and proliferation. Tramp C1 cells were cultured with or without 0.5 × 10−6 m of Cbz for 24 h. Then, DCs were added to uptake ICD induced Tramp C1 cells. After 4 h, CMFDA labeled immunized T cells from donor mice were cocultured with primed DCs. After 5 d of cultivation, Tramp C1 specific T cell proliferation was observed and analyzed by flow cytometry. G) Cbz treatment dependent PD-L1 expression change was investigated after 18 h (PBS, 0.5 × 10−6 m of Cbz).

Article Snippet: After 3 h of culture, 5-chloromethylfluorescein diacetate (CMFDA) dye (BD, USA) labeled T cells were cocultured and the proliferation was tracked for 5 d. The decreased amount of CMFDA fluorescence intensity corresponding to T cell proliferation was analyzed by Flow cytometry (BD, LSRII Fortessa, USA).

Techniques: Over Expression, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, In Vivo, Negative Control, Isolation, Marker, Ex Vivo, Activation Assay, Labeling

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: Microscopic analysis of labeled hBM-MSC. a Vital observation of hBM-MSC stained with Molday ION Rhodamine B™ (Molday), CellTracker™ Green CMFDA (CMFDA), and eGFP (mRNA GFP) over 7 days in vitro culture. *DIV, day in vitro . Scale 50 μm. Measurement of fluorescence signal intensity generated by cells stained with b Molday ION Rhodamine B™ (Molday), c CellTracker™ Green CMFDA (CMFDA), and d transfected with mRNA eGFP on the second and seventh day of in vitro culture. e Comparison of percentage of fluorescent hBM-MSC in all groups. The viability of cells assessed in 7AAD test on the second and seventh day after labeling ( f ). The measurement of relative size ( g ) and granularity (H) of cells. * p < 0.05, ** p < 0.01, *** p < 0.001 ( n = 3–4)

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Labeling, Staining, In Vitro, Fluorescence, Generated, Transfection

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: Phenotypical analysis of hBM-MSC in the 2nd and 7th day of culture after staining with CellTracker™ Green CMFDA (CMFDA), eGFP (mRNA GFP), and Molday ION Rhodamine B™ (Molday) with antibodies directed against proteins: CD90, CD44, SSEA4, and CXCR4 performed by cytometric analysis ( a , c , e , g ). The bar charts ( b , d , f , h ) show the comparison of the percentage of hBM-MSC positive for particular antigen among control, CellTracker™ Green CMFDA (CMFDA), eGFP (mRNA GFP), and Molday ION Rhodamine B™ (Molday)-stained cell groups. * p < 0.05, ** p < 0.01, *** p < 0.001 ( n = 3–4)

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Staining

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: Adipogenesis of hBM-MSC labeled with Molday ION Rhodamine B™ (Molday), CellTracker™ Green CMFDA (CMFDA), and eGFP (mRNA GFP). Adipogenesis occurred in all cell populations ( a ); however, the percentage of differentiating cells ( b ) and persistence of particular dyes ( c ) were significantly different among cell group. * p < 0.05, ** p < 0.01, *** p < 0.001 ( n = 3)

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Labeling

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: Osteogenesis of control, Molday ION Rhodamine B™ (Molday)-labeled, CellTracker™ Green CMFDA (CMFDA)-labeled, and mRNA eGFP (mRNA GFP)-transfected hBM-MSC. All cells were able to undergo osteogenic differentiation ( b ); however, none of them maintained fluorescence ( a )

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Labeling, Transfection, Fluorescence

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: Chondrogenesis of control, Molday ION Rhodamine B™ (Molday)-labeled, CellTracker™ Green CMFDA (CMFDA)-labeled, and mRNA eGFP (mRNA GFP)-transfected hBM-MSC. a A schema shows the developmental stages of micromass creation observed during hBM-MSC osteogenic differentiation. b Table contains data about the presence of particular colony stages during differentiation period in stained hBM-MSC groups. c Pictures of hBM-MSC colonies visible in the fluorescent and light microscope with Alcian blue staining on day 15

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Labeling, Transfection, Staining, Light Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: The real-time PCR analysis of growth factors’ transcript level in cells stained with Molday ION Rhodamine B™ (Molday), CellTracker™ Green CMFDA (CMFDA), and mRNA eGFP (mRNA GFP) in comparison to unlabeled hBM-MSC in the 2nd, 5th, and 7th day after labeling. * p < 0.05, ** p < 0.01, *** p < 0.001 ( n = 5–7)

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Real-time Polymerase Chain Reaction, Staining, Labeling

Journal: Stem Cell Research & Therapy

Article Title: Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity

doi: 10.1186/s13287-019-1296-8

Figure Lengend Snippet: ELISA analysis of GDNF ( a ), IGF-1 ( b ), and HGF ( c ) level in culture media obtained from hBM-MSC stained with Molday ION Rhodamine B™ (Molday), CellTracker™ Green CMFDA (CMFDA), and mRNA eGFP (mRNA GFP) collected on the 2nd, 5th, and 7th day after labeling as compared to unlabeled hBM-MSC. * p < 0.05, ** p < 0.01, *** p < 0.001 ( n = 6)

Article Snippet: hBM-MSCs (Lonza) were cultured at 75-cm 2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Labeling