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Special Issue "Organoid Culture and Characterization Systems for Tissue Engineering Applications"

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biology".

Deadline for manuscript submissions: 30 September 2023 | Viewed by 1771

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Special Issue Editors

Prof. Dr. Manuel Mata

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Guest Editor

Pathology Department, Faculty of Medicine and Odonthology, University of Valencia, Blasco Ibanez Avenue, 17, 46010 Valencia, Spain
Interests: tissue engineering; cell culture; molecular biology; cartilage regeneration; lung cancer

Dr. María Sancho-Tello

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Guest Editor

Pathology Department, Faculty of Medicine and Odonthology, University of Valencia, Blasco Ibanez Avenue, 17, 46010 Valencia, Spain
Interests: human histology; experimental pathology; cell culture; tissue engineering; artificial organs

Special Issue Information

Dear Colleagues,

Recently, the use of organoids is gaining great interest in areas as different from tissue engineering as the manufacture of substitutes for different tissues, like cartilage or skin, or the construction of models for the study of pathophysiological aspects in complex diseases such as cancer or Alzheimer's. The literature shows great heterogeneity in terms of the methodology used for the isolation and/or construction of organoids (using stem cells, human induced pluripotent stem cell (hiPSC) and/or isolated stromal cells), as well as in the different matrices (native decellularized or manufactured) used. The selection of different study parameters (spheroid morphology, matrix generation, angiogenesis, gene expression, etc.) also vary enormously in relation to the final objective of each investigation. Each specific application of organoids has its particular conditions, so it is convenient to have an idea of what is the best approach when generating study systems based on organoids. The objective of this Special Issue is, on the one hand, to define some suitable models for the construction of organoid-based culture systems for specific applications such as, for example, the regeneration of skeletal tissues, the study of cancer or the design of platforms for drug screening for precision medicine approaches. On the other hand, we propose to define which parameters need to be analyzed and by which methodologies, for each specific application.

Prof. Dr. Manuel Mata
Dr. María Sancho-Tello
Guest Editors

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Keywords

organoid manufacture
organoids isolation
3D culture
precision medicine
disease modeling
stem cells
hiPSC
decellularized extracellular matrix

Published Papers (3 papers)

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Research

Open AccessArticle

A Strainer-Based Platform for the Collection and Immunolabeling of Mouse Intestinal Organoids

and

Int. J. Mol. Sci. 2023, 24(17), 13568; https://doi.org/10.3390/ijms241713568 (registering DOI) - 01 Sep 2023

Abstract

Intestinal organoids have emerged as powerful model systems for studying the complex structure and function of the intestine. However, there is a lack of widely applicable methods for the collection, labeling, and imaging of intestinal organoids. In this study, we developed a novel method for loading and labeling intestinal organoids, a method that efficiently collects the organoids and facilitates imaging of their three-dimensional (3D) structure. Based on this strainer platform, mouse intestinal organoids were adequately collected and immobilized, facilitating the immunolabeling workflow to target proteins of the organoids. After evaluation, the strainer size of 40 μm was considered to be more conducive to the collection and labeling of mouse intestinal organoids. More extensive research on organoids of multiple types and species origins will contribute to broadening the applicability of the methodology. Overall, our study proposes an innovative workflow for loading and analyzing intestinal organoids. The combination of a strainer-based collection method, fluorescent labeling, and 3D reconstruction provides valuable insights into the organization and complexity of these tissue models, thereby offering new avenues for investigating intestinal development, disease modeling, and drug discovery. Full article

(This article belongs to the Special Issue Organoid Culture and Characterization Systems for Tissue Engineering Applications)

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Open AccessArticle

Partner, Neighbor, Housekeeper and Dimension: 3D versus 2D Glomerular Co-Cultures Reveal Drawbacks of Currently Used Cell Culture Models

and

Int. J. Mol. Sci. 2023, 24(12), 10384; https://doi.org/10.3390/ijms241210384 - 20 Jun 2023

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Abstract

Signaling-pathway analyses and the investigation of gene responses to different stimuli are usually performed in 2D monocultures. However, within the glomerulus, cells grow in 3D and are involved in direct and paracrine interactions with different glomerular cell types. Thus, the results from 2D monoculture experiments must be taken with caution. We cultured glomerular endothelial cells, podocytes and mesangial cells in 2D/3D monocultures and 2D/3D co-cultures and analyzed cell survival, self-assembly, gene expression, cell–cell interaction, and gene pathways using live/dead assay, time-lapse analysis, bulk-RNA sequencing, qPCR, and immunofluorescence staining. Without any need for scaffolds, 3D glomerular co-cultures self-organized into spheroids. Podocyte- and glomerular endothelial cell-specific markers and the extracellular matrix were increased in 3D co-cultures compared to 2D co-cultures. Housekeeping genes must be chosen wisely, as many genes used for the normalization of gene expression were themselves affected in 3D culture conditions. The transport of podocyte-derived VEGFA to glomerular endothelial cells confirmed intercellular crosstalk in the 3D co-culture models. The enhanced expression of genes important for glomerular function in 3D, compared to 2D, questions the reliability of currently used 2D monocultures. Hence, glomerular 3D co-cultures might be more suitable in the study of intercellular communication, disease modelling and drug screening ex vivo. Full article

(This article belongs to the Special Issue Organoid Culture and Characterization Systems for Tissue Engineering Applications)

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Open AccessArticle

Morphological Characterization of Human Lung Cancer Organoids Cultured in Type I Collagen Hydrogels: A Histological Approach

Irene Monleón-Guinot

Lara Milian

Patricia Martínez-Vallejo

and

Int. J. Mol. Sci. 2023, 24(12), 10131; https://doi.org/10.3390/ijms241210131 - 14 Jun 2023

Viewed by 824

Abstract

The malignity of lung cancer is conditioned by the tumor microenvironment (TME), in which cancer-associated fibroblasts (CAFs) are relevant. In this work, we generated organoids by combining A549 cells with CAFs and normal fibroblasts (NF) isolated from adenocarcinoma tumors. We optimized the conditions for their manufacture in a short time. We evaluated the morphology of organoids using confocal microscopy analysis of F-actin, vimentin and pankeratin. We determined the ultrastructure of the cells in the organoids via transmission electron microscopy and the expression of CDH1, CDH2 and VIM via RT-PCR. The addition of stromal cells induces the self-organization of the organoids, which acquired a bowl morphology, as well as their growth and the generation of cell processes. They also influenced the expression of genes related to epithelial mesenchymal transition (EMT). CAFs potentiated these changes. All cells acquired a characteristic secretory phenotype, with cohesive cells appearing inside the organoids. In the periphery, many cells acquired a migratory phenotype, especially in organoids that incorporated CAFs. The deposit of abundant extracellular matrix could also be observed. The results presented here reinforce the role of CAFs in the progression of lung tumors and could lay the foundation for a useful in vitro pharmacological model. Full article

(This article belongs to the Special Issue Organoid Culture and Characterization Systems for Tissue Engineering Applications)

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Special Issue "Organoid Culture and Characterization Systems for Tissue Engineering Applications"

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