NDLI: In vitro model alveoli from photodegradable microsphere templates.

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In vitro model alveoli from photodegradable microsphere templates.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lewis, Katherine J. R. Tibbitt, Mark W. Zhao, Yi Branchfield, Kelsey Sun, Xin Balasubramaniam, Vivek Anseth, Kristi S.
Description	Country affiliation: United States Author Affiliation: Lewis KJ ( Department of Chemical and Biological Engineering, the BioFrontiers Institute, and the Howard Hughes Medical Institute, University of Colorado at Boulder, 3415 Colorado Ave, 596 UCB, Boulder, CO 80303, USA. kristi.anseth@colorado.edu.)
Abstract	Recreating the 3D cyst-like architecture of the alveolar epithelium in vitro has been challenging to achieve in a controlled fashion with primary lung epithelial cells. Here, we demonstrate model alveoli formed within a tunable synthetic biomaterial platform using photodegradable microspheres as templates to create physiologically relevant, cyst structures. Poly(ethylene glycol) (PEG)-based hydrogels were polymerized in suspension to form microspheres on the order of 120 µm in diameter. The gel chemistry was designed to allow erosion of the microspheres with cytocompatible light doses (≤15 min exposure to 10 mW cm(-2) of 365 nm light) via cleavage of a photolabile nitrobenzyl ether crosslinker. Epithelial cells were incubated with intact microspheres, modified with adhesive peptide sequences to facilitate cellular attachment to and proliferation on the surface. A tumor-derived alveolar epithelial cell line, A549, completely covered the microspheres after only 24 hours, whereas primary mouse alveolar epithelial type II (ATII) cells took â¼3 days. The cell-laden microsphere structures were embedded within a second hydrogel formulation at user defined densities; the microsphere templates were subsequently removed with light to render hollow epithelial cysts that were cultured for an additional 6 days. The resulting primary cysts stained positive for cell-cell junction proteins (ß-catenin and ZO-1), indicating the formation of a functional epithelial layer. Typically, primary ATII cells differentiated in culture to the alveolar epithelial type I (ATI) phenotype; however, each cyst contained â¼1-5 cells that stained positive for an ATII marker (surfactant protein C), which is consistent with ATII cell numbers in native mouse alveoli. This biomaterial-templated alveoli culture system should be useful for future experiments to study lung development and disease progression, and is ideally suited for co-culture experiments where pulmonary fibroblasts or endothelial cells could be presented in the hydrogel surrounding the epithelial cysts.
File Format	HTM / HTML
ISSN	20474830
e-ISSN	20474849
DOI	10.1039/c5bm00034c
Journal	Biomaterials Science
Issue Number	6
Volume Number	3
Language	English
Publisher	Royal Society of Chemistry
Publisher Date	2015-06-01
Publisher Place	Great Britain (UK)
Access Restriction	Open
Subject Keyword	Discipline Biomedical Engineering Discipline Therapeutics Biocompatible Materials Chemistry Cells, Cultured Epithelial Cells Cytology Hydrogels Polyethylene Glycols Pulmonary Alveoli Beta Catenin Metabolism Animals Mice Microspheres Photolysis Research Support, Non-u.s. Gov't Research Support, U.s. Gov't, Non-p.h.s.
Content Type	Text
Resource Type	Article
Subject	Materials Science Biomedical Engineering

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