Automated decellularization of intact, human-sized lungs for tissue engineering.

We developed an automated system that can be used to decellularize whole human-sized organs and have shown lung as an example. Lungs from 20 to 30 kg pigs were excised en bloc with the trachea and decellularized with our established protocol of deionized water, detergents, sodium chloride, and porcine pancreatic DNase. A software program was written to control a valve manifold assembly that we built for selection and timing of decellularization fluid perfusion through the airway and the vasculature. This system was interfaced with a prototypic bioreactor chamber that was connected to another program, from a commercial source, which controlled the volume and flow pressure of fluids. Lung matrix that was decellularized by the automated method was compared to a manual method previously used by us and others. Automation resulted in more consistent acellular matrix preparations as demonstrated by measuring levels of DNA, hydroxyproline (collagen), elastin, laminin, and glycosaminoglycans. It also proved highly beneficial in saving time as the decellularization procedure was reduced from days down to just 24 h. Developing a rapid, controllable, automated system for production of reproducible matrices in a closed system is a major step forward in whole-organ tissue engineering.

[1]  J. Orens,et al.  General overview of lung transplantation and review of organ allocation. , 2009, Proceedings of the American Thoracic Society.

[2]  John P. Soleas,et al.  Evaluation of the structural integrity and extracellular matrix components of tracheal allografts following cyclical decellularization techniques: comparison of three protocols. , 2012, Tissue engineering. Part C, Methods.

[3]  George P McCabe,et al.  Macrophage phenotype and remodeling outcomes in response to biologic scaffolds with and without a cellular component. , 2009, Biomaterials.

[4]  D. Papy-Garcia,et al.  Improved and simple micro assay for sulfated glycosaminoglycans quantification in biological extracts and its use in skin and muscle tissue studies. , 2003, Glycobiology.

[5]  V. Lama.  Update in lung transplantation 2008. , 2009, American journal of respiratory and critical care medicine.

[6]  Daniel J Weiss,et al.  Initial binding and recellularization of decellularized mouse lung scaffolds with bone marrow-derived mesenchymal stromal cells. , 2012, Tissue engineering. Part A.

[7]  Kevin Weiss,et al.  Acellular normal and fibrotic human lung matrices as a culture system for in vitro investigation. , 2012, American journal of respiratory and critical care medicine.

[8]  Zhen W. Zhuang,et al.  Tissue-Engineered Lungs for in Vivo Implantation , 2010, Science.

[9]  Stephen F Badylak,et al.  RETRACTED: Engineered whole organs and complex tissues , 2012, The Lancet.

[10]  Aline M. Betancourt,et al.  A nonhuman primate model of lung regeneration: detergent-mediated decellularization and initial in vitro recellularization with mesenchymal stem cells. , 2012, Tissue engineering. Part A.

[11]  E. Trulock,et al.  The Registry of the International Society for Heart and Lung Transplantation: twenty-first official adult lung and heart-lung transplant report--2004. , 2004, The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation.

[12]  Angela Panoskaltsis-Mortari,et al.  Development of a decellularized lung bioreactor system for bioengineering the lung: the matrix reloaded. , 2010, Tissue engineering. Part A.

[13]  A. Atala,et al.  Decellularization for whole organ bioengineering , 2013, Biomedical materials.

[14]  Stephen F Badylak,et al.  An overview of tissue and whole organ decellularization processes. , 2011, Biomaterials.

[15]  T. Ashikaga,et al.  The effect of age and emphysematous and fibrotic injury on the re-cellularization of de-cellularized lungs. , 2013, Biomaterials.

[16]  Mark F. Lythgoe,et al.  A rat decellularized small bowel scaffold that preserves villus-crypt architecture for intestinal regeneration , 2012, Biomaterials.

[17]  Christian Schuetz,et al.  Regeneration and orthotopic transplantation of a bioartificial lung , 2010, Nature Medicine.

[18]  Jean A. Niles,et al.  Production and assessment of decellularized pig and human lung scaffolds. , 2013, Tissue engineering. Part A.

[19]  T. Jensen,et al.  Can stem cells be used to generate new lungs? Ex vivo lung bioengineering with decellularized whole lung scaffolds , 2013, Respirology.

[20]  Doris A Taylor,et al.  Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart , 2008, Nature Medicine.

[21]  Hiroshi Yagi,et al.  Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix , 2010, Nature Medicine.

[22]  Korkut Uygun,et al.  Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds. , 2011, Annual review of biomedical engineering.

[23]  Anthony Callanan,et al.  Comparison of methods for whole-organ decellularization in tissue engineering of bioartificial organs. , 2013, Tissue engineering. Part B, Reviews.

[24]  M. Estenne,et al.  Bronchiolitis obliterans after human lung transplantation. , 2002, American journal of respiratory and critical care medicine.

[25]  J. Guyette,et al.  Regeneration and Experimental Orthotopic Transplantation of a Bioengineered Kidney , 2013, Nature Medicine.

[26]  Chad A. Cowan,et al.  Generation of multipotent lung and airway progenitors from mouse ESCs and patient-specific cystic fibrosis iPSCs. , 2012, Cell stem cell.

[27]  F. Gage,et al.  A new approach for increasing the organ supply. , 1996, Clinical transplantation.

[28]  Sara Mantero,et al.  Clinical transplantation of a tissue-engineered airway , 2008, The Lancet.