Three-dimensional (3D) tissuelike assemblies (TLAs) of human bronchoepithelial (HBE) cells have been developed for use in in vitro research on infection of humans by respiratory viruses. The 2D monolayer HBE cell cultures heretofore used in such research lack the complex cell structures and interactions characteristic of in vivo tissues and, consequently, do not adequately emulate the infection dynamics of in-vivo microbial adhesion and invasion. In contrast, the 3D HBE TLAs are characterized by more-realistic reproductions of the geometrical and functional complexity, differentiation of cells, cell-to-cell interactions, and cell-to-matrix interactions characteristic of human respiratory epithelia. Hence, the 3D HBE TLAs are expected to make it possible to perform at least some of the research in vitro under more-realistic conditions, without need to infect human subjects.

These Virus Titers indicate rapid growth of virus populations during the first few days.
The TLAs are grown on collagen-coated cyclodextran microbeads under controlled conditions in a nutrient liquid in the simulated microgravitational environment of a bioreactor of the rotating-wall-vessel type. Primary human mesenchymal bronchial-tracheal cells are used as a foundation matrix, while adult human bronchial epithelial immortalized cells are used as the overlying component. The beads become coated with cells, and cells on adjacent beads coalesce into 3D masses. The resulting TLAs have been found to share significant characteristics with in vivo human respiratory epithelia including polarization, tight junctions, desmosomes, and microvilli. The differentiation of the cells in these TLAs into tissues functionally similar to in vivo tissues is confirmed by the presence of tissuelike differentiation marker compounds, including villin, keratins, and specific lung epithelium marker compounds, and by the production of tissue mucin.

In a series of initial infection tests, TLA cultures were inoculated with human respiratory syncytial viruses and parainfluenza type 3 viruses. Infection was confirmed by photomicrographs that showed signs of damage by viruses and virus titers (see figure) that indicated large increases in the populations of viruses during the days following inoculation.

This work was done by Thomas J. Goodwin of Johnson Space Center. For more information, download the Technical Support Package (free white paper) at www.medicaldesignbriefs.com .

This invention is owned by NASA, and a patent application has been filed. Inquiries concerning nonexclusive or exclusive license for its commercial development should be addressed to

the Patent Counsel
Johnson Space Center
(281) 483-0837.

Refer to MSC-24164-1.

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This Brief includes a Technical Support Package (TSP).
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Tissuelike 3D Assemblies of Human Broncho-Epithelial Cells

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Medical Design Briefs Magazine

This article first appeared in the March, 2010 issue of Medical Design Briefs Magazine (Vol. 34 No. 3).

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Overview

The document is a Technical Support Package from NASA, specifically focusing on Tissuelike 3D Assemblies of Human Broncho-Epithelial Cells (MSC-24164-1). It outlines the development and significance of three-dimensional (3D) cell models that replicate the structural and functional characteristics of human respiratory epithelium. These models are essential for studying respiratory diseases and the effects of various pathogens, including viruses.

The introduction emphasizes the importance of creating cell-based models that accurately reflect the complexity and organization of human tissues. Such models are crucial for understanding the innate immune responses of the respiratory system and for developing effective treatments and vaccines against respiratory infections.

The document acknowledges the support received from Wyeth Vaccine Discovery and NASA's Biological Sciences and Applications Division, highlighting the collaborative efforts of various experts in the field. Notable contributions include the expertise of Miguel Suderman in preparing scanning electron microscopy (SEM) and immunohistochemistry (IHC) analyses, as well as guidance from Dr. H.Q. Wang from the University of Texas Medical Branch.

The references section cites several key studies that provide foundational knowledge on the innate host defense mechanisms of the respiratory epithelium, the structural properties of airway epithelium, and the role of epithelial cells in combating respiratory pathogens. These references underscore the scientific context in which the 3D assemblies are developed and their relevance to ongoing research in respiratory health.

The document also includes technical details about the methodologies used in creating these 3D cell models, although specific experimental results and data are not provided in the excerpts. The potential applications of this research extend beyond basic science, suggesting implications for drug development, vaccine testing, and understanding disease mechanisms.

In summary, this Technical Support Package presents a significant advancement in the field of respiratory biology, showcasing how aerospace-related technologies can contribute to biomedical research. The development of Tissuelike 3D Assemblies of Human Broncho-Epithelial Cells represents a promising tool for investigating respiratory diseases and enhancing our understanding of the human immune response in the respiratory tract.