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Pediatr Allergy Immunol. 2017 Dec;28(8):810-817. doi: 10.1111/pai.12810. Epub 2017 Nov 22.

Conditional reprogramming of pediatric airway epithelial cells: A new human model to investigate early-life respiratory disorders.

Wolf S1,2,3, Perez GF1,2,3, Mukharesh L1,2,3, Isaza N1,2,4, Preciado D1,2,5, Freishtat RJ1,2,6, Pillai D1,2,3, Rose MC1,2,3, Nino G1,2,3.

Author information

1
Center for Genetic Research Medicine, Children's National Medical Center, Washington, DC, USA.
2
Department of Pediatrics, George Washington University School of Medicine and Health Sciences, Washington, DC, USA.
3
Division of Pulmonary and Sleep Medicine, Children's National Medical Center, Washington, DC, USA.
4
Division of Neonatology, Children's National Medical Center, Washington, DC, USA.
5
Division of Pediatric Otorhinolaryngology, Children's National Medical Center, Washington, DC, USA.
6
Division of Emergency Medicine, Children's National Medical Center, Washington, DC, USA.

Abstract

BACKGROUND:

Airway epithelial cells (AEC) are quite difficult to access in newborns and infants. It is critically important to develop robust life-extended models to conduct translational studies in this age group. We propose the use of a recently described cell culture technology (conditionally reprogrammed cells-CRC) to generate continuous primary cell cultures from nasal and bronchial AEC of young children.

METHODS:

We collected nasal and/or bronchial AEC from a total of 23 subjects of different ages including newborns/infants/toddlers (0-2 years; N = 9), school-age children (4-11 years; N = 6), and a group of adolescent/adult donors (N = 8). For CRC generation, we used conditioned medium from mitotically inactivated 3T3 fibroblasts and Rho-associated kinase (ROCK) inhibitor (Y-27632). Antiviral immune responses were studied using 25 key antiviral genes and protein production of type III epithelial interferon (IFN λ1) after double-stranded (ds) RNA exposure.

RESULTS:

CRC derived from primary AEC of neonates/infants and young children exhibited: (i) augmented proliferative capacity and life extension, (ii) preserved airway epithelial phenotype after multiple passages, (iii) robust immune responses characterized by the expression of innate antiviral genes and parallel nasal/bronchial production of IFN λ1 after exposure to dsRNA, and (iv) induction of airway epithelial inflammatory and remodeling responses to dsRNA (eg, CXCL8 and MMP9).

CONCLUSION:

Conditional reprogramming of AEC from young children is a feasible and powerful translational approach to investigate early-life airway epithelial immune responses in humans.

KEYWORDS:

airway; bronchial; children; conditionally reprogrammed cells; epithelium; infants; nasal; newborns

PMID:
28981980
PMCID:
PMC5868353
DOI:
10.1111/pai.12810
[Indexed for MEDLINE]
Free PMC Article

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