Characterization of pulmonary cell growth parameters in a continuous perfusion microfluidic environment.
In vitro models of the alveolo-pulmonary barrier consist of microvascular endothelial cells and alveolar epithelial cells cultured on opposing sides of synthetic porous membranes. However, these simple models do not reflect the physiological microenvironment of pulmonary cells, wherein cells are exposed to a complex milieu of mechanical and soluble stimuli. In this report, we studied alveolar epithelial (A549) and microvascular endothelial (HMEC-1) cells within varying microfluidic environments as a first step towards building a microfluidic analog of the gas-exchange interface. We fabricated polydimethylsiloxane (PDMS) microdevices for parallel studies of cell growth under multiple flow rates. Cells adhered and proliferated in the microculture chambers for shear stresses up to approximately 2 x 10(-3) dynes/cm(2), corresponding to media turnover rates of approximately 53 seconds. Proliferation of these cells into confluent monolayers and expression of cell-specific markers (SP-A and CD-31) demonstrated successful pulmonary cell culture in microscale devices, a first for alveolar epithelial cells. These results represent the initial steps towards the development of microfluidic analogs of the alveolo-pulmonary barrier and tissue engineering of the lung.
Nalayanda
Divya D
DD
Division of Pediatric Surgery, Johns Hopkins Medical Institutions, Baltimore, Maryland, USA.
Puleo
Christopher M
CM
Fulton
William B
WB
Wang
Tza-Huei
TH
Abdullah
Fizan
F
D015824:Blood-Air Barrier / Q000737:chemistry / Q000276:immunology / Q000473:pathology*; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004129:Dimethylpolysiloxanes; D042783:Endothelial Cells / Q000166:cytology* / Q000276:immunology; D004847:Epithelial Cells / Q000737:chemistry / Q000473:pathology*; D004867:Equipment Design; D006801:Humans; D008168:Lung / Q000098:blood supply*; D008833:Microcirculation / Q000166:cytology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D010477:Perfusion; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D011650:Pulmonary Alveoli / Q000737:chemistry / Q000473:pathology*; D037662:Pulmonary Surfactant-Associated Protein A / Q000032:analysis; D012828:Silicones; D013314:Stress, Mechanical; D013997:Time Factors
D004129:Dimethylpolysiloxanes; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D037662:Pulmonary Surfactant-Associated Protein A; D012828:Silicones; C013830:baysilon