A Two-Dimensional Human Minilung System (Model) for Respiratory Syncytial Virus Infections.

Esmeralda Magro-Lopez,Alberto Zambrano,Maria Martin-Vicente,Trinidad Guijarro,Isabel Liste,Isidoro Martinez

doi:10.3390/v9120379

Abstract

Human respiratory syncytial virus (HRSV) is a major cause of serious pediatric respiratory diseases that lacks effective vaccine or specific therapeutics. Although our understanding about HRSV biology has dramatically increased during the last decades, the need for adequate models of HRSV infection is compelling. We have generated a two-dimensional minilung from human embryonic stem cells (hESCs). The differentiation protocol yielded at least six types of lung and airway cells, although it is biased toward the generation of distal cells. We show evidence of HRSV replication in lung cells, and the induction of innate and proinflammatory responses, thus supporting its use as a model for the study of HRSV–host interactions.

Highlights

Functional Unit for Research into Chronic Diseases (UFIEC), Institute of Health Carlos III, 28220 Madrid, Spanish National Center for Microbiology (CNM), Institute of Health Carlos III, 28220 Madrid, Spain; These authors contributed to this work
A549, or primary epithelial cells derived from human lung airways
It was followed by an outflow of a series of Transwell systems of well-differentiated primary airway epithelial cell cultures (WD-PAECs) displaying a pseudostratified epithelium comprising basal, globet, and ciliated cells [4,5,6,7]

Summary

Introduction

Chronogram of the differentiation strategy followed to generate the two-dimensional minilung from hESCs (human embryonic stem cells). Day hESCs: human embryonic stem cells; MEFs: mouse embryonic fibroblasts; BMP4: bone morphogenic protein 4; ROCK: rho-associated coiled-coil containing protein kinase; ACVR: activin A receptor; FGFR: fibroblast growth factor receptor; TGF: tumor growth factor; IWP2: WNT inhibitor; CHIR99021: WNT activator; KGF: keratinocyte growth factor; FGF: fibroblast growth factor; EGF: epithelial growth factor; IBMX: 3-isobutyl-1-methylxanthine; cAMP: 30 ,50 cyclic adenosine monophosphate As reported before [10], we confirmed the expression of markers of terminal differentiation around day 50 (data not shown) [12] a time from which the amount of mature epithelial cells gradually increases.

Results

Conclusion