In absence of an in vivo model to study diseases associated with lung damage, we propose engineering a functional lung organ in vitro. A 3D lung scaffold was generated using a mouse lung that was decellularized with detergents. The lung scaffolds were recellularized using a bioreactor consisting of a peristaltic pump that periodically pushed media and cells into the scaffold to recapitulate lung respiration. To direct the mouse embryonic stem cells (mESC) differentiation toward the endoderm lineage, the media was supplemented with lung homogenate and Activin A. qRT-PCR demonstrated that, compared to the mESC control, there was an upregulation of gene expression in the heterogeneous cell population indicating differentiation. Haematoxylin and Eosin staining of lung tissue demonstrated positive recellularization and was further confirmed by immunohistochemistry. This study provides valuable insights on how scaffolds and media influences cell culture differentiation of vital tissues to study tissue damage and injury repair.
In Vitro Generation of Lung Tissue using a 3D Scaffold Matrix
Advisor: Dr. Tzvia Abramson
Author: Melad Askndafi