Lung bioscaffolds: comparative lung decellularization techniques

Abstract

While the final treatment for end‐stage lung failure remains lung transplantation, less than 20% of donor lungs are considered suitable for transplantation. An innovation to improve the donor lung pool supply involves decellularization of lungs to provide an acellular three‐dimensional bioscaffold that can be reseeded with lung specific progenitor cells. With no existing gold standard method for lung decellularization to date, it is important to establish an efficient and effective method that removes all DNA and intracellular structural proteins. These techniques also involve testing the bioscaffold integrity, and instituting a cryopreservation method such that a lung bioscaffold repository can be created.Rat lungs were decellularized using static and continuous perfusion decellularization protocols. Static decellularization was done using a 24 and 72‐hour protocol, and continuous decellularization was done using a 75 hour protocol. Triton X‐100, Sodium Deoxycholate or Sodium Dodecyl Sulfate, NaCl, and porcine pancreatic DNAase was used to decellularize the lungs, with the variation being a difference in incubation times. Differences between techniques were determined by histological analysis via H&E and DAPI stains evaluating nuclei and structural proteins. Bioscaffold integrity tests following both decellularization and cryopreservation were also performed.Grant Funding Source: N/A