Development of perfusion bioreactor for whole organ engineering — a culture system that enhances cellular engraftment, survival and phenotype of repopulated pancreas

Abstract

Whole organ engineering has emerged as a promising alternative avenue to fill the gap of donor organ shortage in organ transplantation. Recent breakthroughs in the decellularization of solid organs and repopulation with desired cell populations have generated neo-organ constructs with promising functional outcomes. The realization of this goal requires engineering advancement in the perfusion-based bioreactors to (i) efficiently deliver decellularization agents, followed by (ii) its reconstruction with relevant cell types and (iii) maintenance of viability and function of the repopulated organ. In this study, we report the development and assembly of a perfusion bioreactor with the potential to enable regenerative reconstruction of pancreas. The assembled bioreactor is versatile to efficiently decellularize multiple organs, as demonstrated by complete decellularization of pancreas, liver and heart in the same set-up. Further, the same system is amenable to support organ repopulation with diverse cell types. Using our in-house bioreactor system, we demonstrate pancreas repopulation with both immortalized MIN-6 beta cells and differentiating human pluripotent stem cells. Importantly, we show the significant advantage of perfusion culture over static culture in enhancing cell engraftment, viability and phenotypic maintenance of the repopulated pancreas. In addition, this study is a significant step forward for whole organ engineering as it will facilitate cost-effective and easy assembly of perfusion bioreactors to enable rapid advancement in regenerative organ reconstruction.

Decellularized extracellular matrix scaffolds: Recent trends and emerging strategies in tissue engineering

“…Perfusion is a typical decellularized route for larger, thicker tissues or whole organs, which refers to cannulating organs/tissues and subsequently establishing a channel for circulating agents through their intrinsic vascular system [ 47 ]. The efficiency of perfusion is primarily determined by perfusion route (artery or portal vein route), infusion mode (antegrade or retrograde), perfusion paraments (pressure, temperature, and flow rate), types and concentrations of perfusate, and the dimension of organ/tissue [ [48] , [49] , [50] , [51] , [52] , [53] ].…”

Functional acellular matrix for tissue repair

“…Perfusion is a typical physical decellularization method that involves the cannulation of an organ/tissue followed by the formation of a channel for the flow of a circulating agent through the intrinsic vascular system [ 100 ]. For large thick tissues and whole organs, this method minimizes ECM damage inflicted by excessive forces while maximizing decellularization deep inside the organ [ 5 ].…”

Towards human organ generation using interspecies blastocyst complementation: Challenges and perspectives for therapy

“…Decellularized scaffolds have been made for various organs, including the lung, heart, kidney, liver, intestine, bladder, corneas, limbs, pancreas, and vasculature. ( Ott et al, 2008 ; Hashimoto et al, 2010 ; Petersen et al, 2010 ; Yang et al, 2010 ; Quint et al, 2011 ; Totonelli et al, 2012 ; Song et al, 2013a ; Gerli et al, 2018 ; Goh et al, 2019 ).…”