Alveolar epithelial differentiation of human induced pluripotent stem cells in a rotating bioreactor
| Autor: | Julio J. Mendez, Laura E. Niklason, Peter F. Bove, Amogh Sivarapatna, Micha Sam Brickman Raredon, Mahboobe Ghaedi |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Cellular differentiation
Alveolar Epithelium Population Immunocytochemistry Caveolin 1 Induced Pluripotent Stem Cells Biophysics Bioengineering Biology Article Flow cytometry Biomaterials Bioreactors Tissue engineering medicine Humans education Induced pluripotent stem cell Cells Cultured education.field_of_study Membrane Glycoproteins medicine.diagnostic_test Type-II Pneumocytes Membrane Proteins Cell Differentiation Epithelial Cells respiratory system Cell biology Aquaporin 5 Pulmonary Alveoli Mechanics of Materials Ceramics and Composites |
| Popis: | Traditional stem cell differentiation protocols make use of a variety of cytokines including growth factors (GFs) and inhibitors in an effort to provide appropriate signals for tissue specific differentiation. In this study, iPSC-derived type II pneumocytes (iPSC-ATII) as well as native isolated human type II pneumocytes (hATII) were differentiated toward a type I phenotype using a unique air–liquid interface (ALI) system that relies on a rotating apparatus that mimics in vivo respiratory conditions. A relatively homogenous population of alveolar type II-like cells from iPSC was first generated (iPSC-ATII cells), which had phenotypic properties similar to mature human alveolar type II cells. iPSC-ATII cells were then cultured in a specially designed rotating culture apparatus. The effectiveness of the ALI bioreactor was compared with the effectiveness of small molecule-based differentiation of type II pneumocytes toward type 1 pneumocytes. The dynamics of differentiation were examined by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), flow cytometry and immunocytochemistry. iPSC-ATII and hATII cells cultured in the ALI bioreactor had higher levels of type I markers, including aquaporin-5(AQ5), caveolin-1, and T1α, at both the RNA and protein levels as compared with the flask-grown iPSC-ATII and hATII that had been treated with small molecules to induce differentiation. In summary, this study demonstrates that a rotating bioreactor culture system that provides an air–liquid interface is a potent inducer of type I epithelial differentiation for both iPS-ATII cells and hATII cells, and provides a method for large-scale production of alveolar epithelium for tissue engineering and drug discovery. |
| Databáze: | OpenAIRE |
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