Conditionally reprogrammed asthmatic bronchial epithelial cells express lower <i>FOXJ1</i> at terminal differentiation and lower <i>IFNs</i> following RV-A1 infection.
Punnam Chander VeeratiKristy S NicholJane M ReadNathan W BartlettPeter A B WarkDarryl A KnightChristopher L GraingeAndrew T ReidPublished in: American journal of physiology. Lung cellular and molecular physiology (2022)
Primary bronchial epithelial cells (pBECs) obtained from donors have limited proliferation capacity. Recently, conditional reprogramming (CR) technique has overcome this and has provided the potential for extended passaging and subsequent differentiation of cells at air-liquid interface (ALI). However, there has been no donor-specific comparison of cell morphology, baseline gene expression, barrier function, and antiviral responses compared with their "parent" pBECs, especially cells obtained from donors with asthma. We, therefore, collected and differentiated pBECs at ALI from mild donors with asthma (<i>n</i> = 6) for the parent group. The same cells were conditionally reprogrammed and later differentiated at ALI. Barrier function was measured during the differentiation phase. Morphology and baseline gene expression were compared at terminal differentiation. Viral replication kinetics and antiviral responses were assessed following rhinovirus (RV) infection over 96 h. Barrier function during the differentiation phase and cell structural morphology at terminal differentiation appear similar in both parent and CR groups, however, there were elongated cell structures superficial to basal cells and significantly lower <i>FOXJ1</i> expression in CR group. <i>IFN</i> gene expression was also significantly lower in CR group compared with parent asthma group following RV infection. The CR technique is a beneficial tool to proliferate pBECs over extended passages. Considering lower <i>FOXJ1</i> expression, viral replication kinetics and antiviral responses, a cautious approach should be taken while choosing CR cells for experiments. In addition, as lab-to-lab cell culture techniques vary, the most appropriate technique must be utilized to best match individual cell functions and morphologies to address specific research questions and experimental reproducibility across the labs.
Keyphrases
- gene expression
- induced apoptosis
- cell cycle arrest
- mycobacterium tuberculosis
- single cell
- dna methylation
- chronic obstructive pulmonary disease
- endoplasmic reticulum stress
- poor prognosis
- lung function
- sars cov
- cell death
- oxidative stress
- immune response
- dendritic cells
- risk assessment
- climate change
- binding protein
- cystic fibrosis
- cell proliferation
- long non coding rna