Abstract
Rationale: The airways in chronic obstructive pulmonary disease (COPD) are characterized by inflammation, goblet cell hyperplasia, decreased ciliated cell numbers and airway remodeling with an altered composition/deposition of the extracellular matrix (ECM). We aimed to develop a lung ECM hydrogel-based spatiotemporal model to investigate bidirectional interactions with fibroblasts and epithelial cells in relation to ECM changes.
Methods: Porcine lungs were decellularized, lyophilized, ground into a powder, porcine pepsin-solubilized for 48h, buffered with PBS, pH neutralized and gelated at 37°C for 30min. Non-diseased primary lung fibroblasts (250,000 per ml) were mixed with hydrogels in transwells. After 24h the hydrogels were coated with Matrigel®, Collagen type IV or left uncoated. After 24h, 90,000 non-diseased primary bronchial epithelial cells (PBEC) were seeded on top. The hydrogels were imaged using light microscopy, stained for live and dead cells and paraffin embedded for histology.
Results: The fibroblasts inside the ECM hydrogels were viable for up to 5 days, displaying a spindle-shaped morphology. The Matrigel and Collagen IV formed a thin layer on top of the ECM hydrogel which was visualized by a collagen IV immunohistochemical staining. The PBECs grew to a confluent layer in 3-5 days on top of the coated and non-coated ECM hydrogels with no apparent difference in the PBEC layer between them.
Conclusions: The lung ECM hydrogel in combination with a basement membrane like coating allowed for the co-culture of primary lung fibroblasts and PBECs in their correct dimensionality resembling the airway in vivo. The next step is to expose the apical surface of the PBEC layer to air to stimulate the development of a mucociliated epithelial layer.
Footnotes
Cite this article as ERJ Open Research 2021; 7: Suppl. 6, 110.
This is an ERS Lung Science Conference abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).
- Copyright ©the authors 2021