Characterisation of a novel 3D model of the airways

Asthma and COPD are major causes of mortality and morbidity worldwide. Whilst the aetiology underlying both diseases are different, chronic inflammation and structural changes to the airways, termed remodelling, (including mucous hypersecretion, fibrosis and airway wall thickening) is observed in both diseases. We have developed a novel 3D cell culture model (bronchospheres) of the airways using p63+/Krt6+ primary human basal epithelial cells. Bronchospheres capture more of the relevant complexity of the airways when compared to 2D cultures such as air liquid interface (ALI) cultures. Normal Human Epithelial (NHBEC), Asthmatic Human Epithelial (AHBEC) and COPD Human Epithelial (CHBEC) cells were obtained from Lonza and seeded onto matrigel at Passage 2. Basal cells were cultured for 14-18 days being fed on day 2 and day 8. Basal cell to bronchosphere transformation was visualised using light and video microscopy. NHBEC, AHBEC and CHBEC formed bronchospheres developing a fully functional pseudostratified epithelial structure with an inner lumen, mucus secreting goblet cells and beating cilia. There are differences between the rate at, which basal cells from different disease states form bronchospheres (NHBEC= 18 days, AHBEC= 20 days and CHBEC = 24 days) and the average size that they reach (Figure 1). The physiology of bronchospheres make them ideal intermediates between 2D and in vivo models.