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

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.

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

“…At present, nonspecific staining caused by strainer materials is acceptable until new strainer materials are developed. In addition, for organoid immune labeling, imaging observation without removal of gels may be achieved in type I collagen gels [38]. Compared with the strainer platform, direct immune labeling of type I collagen gels may remove a large number of organoids during execution.…”

Advanced lung organoids and lung-on-a-chip for cancer research and drug evaluation: a review

“…In 2023, Monleon-Guinot et al used transmission electron microscopy to observe and compare the morphology of two groups of organoid models: A549 mixed cultures with CAFs and A549 mixed cultures with normal fibroblasts (NF). They found that CAFs promote the upregulation of the expression levels of EMT-related genes such as CDH1 and VIM (Monleon-Guinot et al, 2023). These experimental results indicate that novel 3D models can demonstrate the complex interaction of cancer cell proliferation, invasion, and metastasis with CAFs (Kalluri and Zeisberg, 2006), providing a new platform for tumour mechanism research and drug evaluation.…”

The Nexus of Inflammation-Induced Epithelial-Mesenchymal Transition and Lung Cancer Progression: A Roadmap to Pentacyclic Triterpenoid-Based Therapies

“…Collagens play an integrative role in both normal and tumor tissues, connecting different cell types [83,84]. In addition to their structural function, collagens I and XVII have been found to be induced during inflammation and induce EMT in lung cancer cells [85][86][87][88].…”