Culture of iPSCs Derived Pancreatic β-Like Cells In Vitro Using Decellularized Pancreatic Scaffolds: A Preliminary Trial

Abstract

Diabetes mellitus is a disease which has affected 415 million patients in 2015. In an effort to replace the significant demands on transplantation and morbidity associated with transplantation, the production of β-like cells differentiated from induced pluripotent stem cells (iPSCs) was evaluated. This approach is associated with promising decellularized scaffolds with natural extracellular matrix (ECM) and ideal cubic environment that will promote cell growth in vivo. Our efforts focused on combining decellularized rat pancreatic scaffolds with mouse GFP+-iPSCs-derived pancreatic β-like cells, to evaluate whether decellularized scaffolds could facilitate the growth and function of β-like cells. β-like cells were differentiated from GFP+-iPSCs and evaluated via cultivating in the dynamic circulation perfusion device. Our results demonstrated that decellularized pancreatic scaffolds display favorable biochemical properties. Furthermore, not only could the scaffolds support the survival of β-like cells, but they also accelerated the expression of the insulin as compared to plate-based cell culture. In conclusion, these results suggest that decellularized pancreatic scaffolds could provide a suitable platform for cellular activities of β-like cells including survival and insulin secretion. This study provides preliminary support for regenerating insulin-secreting organs from the decellularized scaffolds combined with iPSCs derived β-like cells as a potential clinical application.

Strategies based on organ decellularization and recellularization

“…In a recently published study from Wan et al, decellularized pancreatic tissues were recellularized with mouse iPSC-derived ß-like cells and cultured for 5 days. Furthermore, the authors showed increased expression of the insulin gene compared to a 2D culture [147]. An as yet neglected point is the importance of the islet niche for recellularization.…”

“…Only a few publications have examined the decellularization of whole pancreas and the subsequent repopulation with islets of Langerhans. Protocols for perfusion‐based decellularization of the mouse, rat, pig and human pancreas have been published , but use different detergents and times to decellularize the pancreas. Detergents include nonionic tensides such as Triton X‐100 and enzymatic agents such as DNAse.…”

Using GRGDSPC peptides to improve re‐endothelialization of decellularized pancreatic scaffolds

“…Not only do they provide space for cell growth but they also promote cell migration, differentiation, and metabolism through interactions with the cell matrix . In our previous study, we succeeded in recellularizing the decellularized pancreatic scaffold with induced pluripotent stem cell‐derived pancreatic β‐like cells . Unfortunately, vascularization of the engineered tissue remains the main challenge for making engineered pancreatic tissue.…”

“…22 In our previous study, we succeeded in recellularizing the decellularized pancreatic scaffold with induced pluripotent stem cell-derived pancreatic β-like cells. 23 Unfortunately, vascularization of the engineered tissue remains the main challenge for making engineered pancreatic tissue. It has been shown that the exposed ECM in the vascular network is responsible for platelet activation and formation of a thrombus.…”

Bioengineering the Pancreas: Cell-on-Scaffold Technology

“…Thus stem cells can be considered a potent and encouraging cell source for tissue engineering [124]. Accordingly to this concept, Wan et al has just proposed a study about the possibility to culturing iPSCs derived pancreatic β cells on decellularized ECM [125]. After peristaltic artery perfusion with detergents, decellularized ECM-based pancreas were seeded with already differentiated into β cells like iPSCs (approx.…”