(191x) Construction of Human Bronchial Epithelium Culture Platform for Inhalation Drug Development

The culture of human bronchial epithelium is widely used to enable the in vitro research to understand the development of lung diseases and to facilitate the development of suitable treatment. Although many standard procedures for the culture of a bronchial epithelial cell line are readily available using transwell inserts, the culture steps are considered to be labor intensive and the cell conditions are varied with the handling (e.g. the remained medium in the apical side, the interval of mucus washing). In this work, we demonstrate a semi-automated platform for long-term mucociliary epithelium culture allowing the users to maintain the cell culture with minimum intervention. This highly integrated microfluidic-based platform was fabricated using a rapid prototyping technique based on laser patterned polymeric membranes/films/sheets and lamination. Microfluidic channels were patterned using laser cut acrylic, polycarbonate and PET sheets. Each layer was laminated using biocompatible silicone based adhesive transfer tapes to form the highly integrated bioreactor.

The platform mainly combines three parts: a cell culture device, a multi-functional reservoir, and a flow management system including a customized peristaltic pump, pinch valves, and fluid network. The cell culture steps can be simply controlled by switching the operation modes allowing: (1) cell seeding; (2) cell proliferation under liquid-liquid interface; (3) cell differentiation under air-liquid interface; (4) repeatedly mucus washing. The results demonstrate that the human airway epithelial cell (Calu-3) can be cultured on the platform with a high viability. The dynamic cell culture condition can simulate a physiologic environment that is capable of maintaining cell differentiation and tissue-specific functions. Finally, by introducing the aerosolized drug in the bronchial epithelium platform using a nebulizer, the adsorption ability of drug can be verified by measuring the concentration of dissolved drug in basolateral side of the tissue. In addition to bronchial tissue culture, the versatile combination of culture steps makes the tissue culture platform suitable to adapt to other in vitro human tissue culture models.