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Visualizing Hepatic Immunity through the Eyes of Intravital Microscopy
Published in Margarida M. Barroso, Xavier Intes, In Vivo, 2020
Maria Alice Freitas-Lopes, Maísa Mota Antunes, Raquel Carvalho-Gontijo, Érika de Carvalho, Gustavo Batista Menezes
Many immune cell markers are named following the clusters of differentiation (CD) nomenclature, which aims to provide targets for cell immunophenotyping. The majority of these surface antigens is not only specific to a unique cell type. A classic example is the CD11b, an integrin family member, expressed on the surface of many leukocytes including monocytes, neutrophils, NK cells, granulocytes, and macrophages. To better identify a specific cell type, it is important to search for a marker expressed exclusively for this lineage or cell. Additionally, imaging experiments allow the evaluation of morphological aspects of the cells, as well as the size and presence of dendrites, for example, which facilitates their identification jointly to the marking.
Using co-axial electrospray deposition to eliminate burst release of simvastatin from microparticles and to enhance induced osteogenesis
Published in Journal of Biomaterials Science, Polymer Edition, 2019
Xiaowei Yuan, Mei Zhang, Yilong Wang, He Zhao, Dahui Sun
SD rat BMSCs were used to investigate the in vitro cytocompatibility of the electrosprayed MPs. BMSCs were isolated based on a method modified from that of Zhu et al. [42]. In brief, four 120 g male SD rats were euthanized by cervical dislocation. After the rats were disinfected, the soft tissues, such as muscles and tendons, were carefully disassociated completely from the tibias and femurs using dissecting scissors and a scalpel, to avoid contamination. Bone marrow cavities of the femurs and tibias were slowly flushed with culture media under sterile conditions until the bones become pale. The cells were seeded at 100 cells per cm2 and grown in a 90-mm sterile culture dish with complete culture medium (L-DMEM supplemented with 10% FBS and 1% penicillin-streptomycin solution) at 37 °C with 5% CO2 in a humidified incubator. All samples were processed within 30 min of the animal death to ensure high stem cell viability. The growth medium was changed every two or three days. Cells were subcultured at a split ratio of 1:3 (resuspended in 75 cm2 cell culture flask [Corning, NY, USA]) by trypsin-EDTA solution when they reached approximately 80% confluence. While non-adhesive cells were removed by replacing the media with fresh media, the adhesive BMSCs were cultured to the third-generation. The expression of surface antigens was detected using flow cytometry (Abcam, Cambridge, UK) with CD11b, CD29, CD45 and CD90. BMSCs were plated in triplicate and maintained in adipogenic and osteogenic induction medium for 21 days for adipogenic differentiation and osteogenic differentiation, respectively. The undifferentiated cells were used as controls.