generating-free-floating-normal-human-epithelial-fibroblast-spheroid

Generating Free-floating Normal Human Epithelial-Fibroblast Spheroid Co-Cultures

Three-dimensional (3D) culture systems have become increasingly popular due to their ability to reproduce natural cell properties and architectures, thus ...

University of Duisburg-Essen

Organoids, as near-physiological 3D culture systems, offer new opportunities to study the pathogenesis of various organs in mimicking the cellular complexity and functionality of human organs.

Matrix-free human lung organoids derived from induced pluripotent stem cells to model lung injury.

The cellular responses of classical 2D flat monolayer cell culture systems provide only very limited reliable predictions about possible outcomes of corresponding animal experiments and clinical studies, which is due, among other things, to the lack of (bi)directional signaling transmission between different cell types and the lack of a structural microenvironment. To study the interactive communication between different cell types in vitro, two main co-culture methods have emerged as central techniques. In the indirect co-culture method, different cells are cultured physically separately (e.g., using transwell inserts) but can communicate with each other via secreted factors (paracrine mechanism). In the direct co-culture method, the different cells have direct physical contact, which enables direct interactions. Regarding the latter method, the cultivation of cells as spherical cell aggregates, so-called spheroids, embedded in a semi-solid extracellular matrix has been established as an in vivo-related, more complex cell culture model with different functional cell states according to cell-cell and cell-ECM interactions as well as oxygen and nutrient gradients. Here, we present a matrix-free method for direct spheroidal co-cultivation of human bronchial epithelial cells and fibroblasts, which can be considered as an in vivo-approximated cultivation method, especially with regard to the cellular composition of the respective spheroids.

Matrix-Free Normal Human Epithelial-Fibroblast 3D Spheroid Cultures for In Vitro Lung Modeling.

Lung diseases are one of the leading causes of death worldwide, and the global burden of these respiratory diseases continues to increase. Therefore, there is a need for accurate models for basic and translational research. In addition to animal models, the development of alternative in vitro model systems is progressing rapidly, ranging from advanced lung cell cultures to complex tissue-engineered lungs. Human lung organoids have become easily transferable three-dimensional in vitro model systems for lung disease modeling. Here, we present a detailed protocol for a rather simple and therefore very practical but reliable method to generate lung organoids from induced pluripotent stem cells (iPSCs) without relying on a matrix, which would represent a step forward toward animal-origin and/or component-free in vitro modeling.

Article	Year
Matrix-free human lung organoids derived from induced pluripotent stem cells to model lung injury. Stem cell research & therapy\| PubMed ID: 39696649	2024
Matrix-Free Normal Human Epithelial-Fibroblast 3D Spheroid Cultures for In Vitro Lung Modeling. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 40106147	2025
Free-Floating Human Lung Organoids Derived from Induced Pluripotent Stem Cells. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 40106148	2025

Lisa Marie Stasch

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