Recellularization of decellularized lung scaffolds is enhanced by dynamic suspension culture.

Aurélie Crabbé,Shameema F Sarker,Daniel J Weiss,Zachary D Borg,Cheryl A Nickerson,Jennifer Barrila,James J Lee,Nicholas R Bonenfant,Yulong Liu,Monica Soncini

doi:10.1371/journal.pone.0126846

Aurélie Crabbé, Shameema F Sarker + Show 8 more

Open Access

https://doi.org/10.1371/journal.pone.0126846

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Abstract

Strategies are needed to improve repopulation of decellularized lung scaffolds with stromal and functional epithelial cells. We demonstrate that decellularized mouse lungs recellularized in a dynamic low fluid shear suspension bioreactor, termed the rotating wall vessel (RWV), contained more cells with decreased apoptosis, increased proliferation and enhanced levels of total RNA compared to static recellularization conditions. These results were observed with two relevant mouse cell types: bone marrow-derived mesenchymal stromal (stem) cells (MSCs) and alveolar type II cells (C10). In addition, MSCs cultured in decellularized lungs under static but not bioreactor conditions formed multilayered aggregates. Gene expression and immunohistochemical analyses suggested differentiation of MSCs into collagen I-producing fibroblast-like cells in the bioreactor, indicating enhanced potential for remodeling of the decellularized scaffold matrix. In conclusion, dynamic suspension culture is promising for enhancing repopulation of decellularized lungs, and could contribute to remodeling the extracellular matrix of the scaffolds with subsequent effects on differentiation and functionality of inoculated cells.

Highlights

Chronic obstructive pulmonary disease (COPD) affects over 64 million people worldwide and is predicted by the World Health Organization to become the third leading cause of mortality by 2030
While this study demonstrated enhanced cell differentiation and viability of embryonic stem cells (ESC) on decellularized scaffolds compared to other extracellular matrix (ECM) scaffolds in the rotating wall vessel (RWV), the authors focused on the role of different ECM scaffolds as identical static controls were not included
Decellularized lung scaffolds recellularized with mesenchymal stromal (stem) cells (MSCs) for a total of 14 days had more abundant cells in the bioreactor as compared to static conditions (Fig 2AD versus Fig 2Ad)

Summary

Introduction

Chronic obstructive pulmonary disease (COPD) affects over 64 million people worldwide and is predicted by the World Health Organization to become the third leading cause of mortality by 2030. While allogeneic lung transplantation is the only definitive treatment for the growing number of patients with end-stage lung disease, only one out of four patients on the organ waiting list undergoes transplantation, given the limited availability of donor organs. The clinical success of lung transplantation is hampered by lifelong immunosuppression and chronic rejection, reflected in a 10–20% survival rate 10 years post-transplantation [1].

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