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A 3D Cell-Culture System That Uses Nano-Fibrillated Bacterial Cellulose to Prepare a Spherical Formulation of Culture Cells.

Biol Pharm Bull

January 2025

Department of Pharmacokinetics and Biopharmaceutics, Institute of Biomedical Sciences, Tokushima University, Tokushima 770-8505, Japan.

A 3-dimensional (3D) cell culture is now being actively pursued to accomplish the in vivo-like cellular morphology and biological functions in cell culture. We recently obtained nano-fibrillated bacterial cellulose (NFBC). In this study, we developed a novel NFBC-based 3D cell-culture system, the OnGel method, and the Suspension method.

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The epithelial and mesenchymal features of colorectal adenocarcinoma (CRAC) cell lines were compared in two-dimensional (2D) and three-dimensional (3D) cultures. In 2D cultures, the three CRAC cell lines exhibited epithelial characteristics with high E-cadherin and low vimentin levels, whereas two exhibited mesenchymal traits with opposite expression patterns. In 3D cultures using low-attachment plates, mesenchymal cells from 2D cultures showed reduced vimentin mRNA levels.

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A thermoplastic chip for 2D and 3D correlative assays combining screening and high-resolution imaging of immune cell responses.

Cell Rep Methods

January 2025

Department of Applied Physics, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden; Department of Medicine, Center for Infectious Medicine, Huddinge, Karolinska Institutet, Stockholm, Sweden. Electronic address:

We present an easy-to-use, disposable, thermoplastic microwell chip designed to support screening and high-resolution imaging of single-cell behavior in two- and three-dimensional (2D and 3D) cell cultures. We show that the chip has excellent optical properties and provide simple protocols for efficient long-term cell culture of suspension and adherent cells, the latter grown either as monolayers or as hundreds of single, uniformly sized spheroids. We then demonstrate the applicability of the system for single-cell analysis by correlating the dynamic cytotoxic response of single immune cells grown under different metabolic conditions to their intracellular cytolytic load at the end of the assay.

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In recent years, three-dimensional (3D) cultures of tumor cells has emerged as an important tool in cancer research. The significance of 3D cultures, such as tumor spheroids, lies in their ability to mimic the in vivo tumor microenvironment more precisely, offering a nuanced understanding of immune responses within the context of tumor progression. In fact, the infiltration of cytotoxic lymphocytes is key to determining patients' prognosis in several types of cancer and response to immunotherapy.

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The emerging field of 3D organ modeling encounters several imaging issues in particular related to antigen retrieval and sample loss during staining processes. Due to their compact shape, several antibodies fail to penetrate intact organoids or spheroids. Histology of organoids can be approached by paraffin inclusion and sectioning at 5 μm as performed for biopsies.

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