3D Culture of MIN-6 Cells on Decellularized Pancreatic Scaffold: In Vitro and In Vivo Study

Abstract

Type 1 diabetes is an autoimmune disease which is due to the lack of β cells. The ideal therapy to cure the disease is pancreas transplantation, but its application is confined to a limited number of people due to the shortage of organ and the need for life-long immunosuppression. Regenerative medicine methods such as a tissue engineered pancreas seem to provide a useful method. In order to construct a microenvironment similar to the native pancreas that is suitable for not only cell growth but also cellular function exertion, a decellularized mouse pancreas was used as a natural 3D scaffold in this experiment. MIN-6 β cells were planted in the bioscaffold. The cell engraftment was verified by HE staining and SEM. Immunostaining procedures were performed to confirm the normal function of the engrafted cells. qRT-PCR demonstrated that insulin gene expression of the recellularized pancreas was upregulated compared with conventional plate-cultured cells. In vivo experiment was also accomplished to further evaluate the function of the recellularized bioscaffold and the result was inspiring. And beyond doubt this will bring new hope for type 1 diabetic patients.

Decellularized Pancreas Matrix Scaffolds for Tissue Engineering Using Ductal or Arterial Catheterization

“…The absence of any nuclear material was very promising, as remnants of DNA fragments in a decellularized scaffold can cause cytocompatibility in vitro and immunological responses after implantation [Brown et al, 2009;Nagata et al, 2010]. In a recent study, a pancreatic bioscaffold was prepared using Triton/ammonium hydroxide perfused through the hepatic portal vein and recellularized with MIN-6 and β-like cells [Wu et al, 2015a]. In another research, a mouse pancreatic bioscaffold was produced based on SDS/Triton perfused through the anterior hepatic portal vein and considered to be capable of mimicking the natural pancreas for pancreatic tissue engineering [Goh et al, 2013].…”

“…Indeed, ECM proteins not oxygen and nutrients, for example, to seeded cells [Wu et al, 2015a]. Methylene blue injection revealed slow flow into the ductal and vascular networks of decellularized pancreas.…”

Tissue and organ decellularization in regenerative medicine

“…The first perfused decellularization of pancreas was carried out in 2009 with a porcine organ, and then applied in mouse . The decellularized tissues were used to seed cells and islets and their in vitro functions were studied as well as the biocompatibility of these bioartificial pancreata was investigated in vivo . Later, rat pancreata were decellularized and a new method was introduced to infuse islets into the ducts, veins and arteries .…”

“…It is very important for scaffolds made from decellularized tissues and organs to have certain mechanical properties matching the site of implantation of the tissue or organ to be substituted. Creating less damage to the tissue during decellularization could yield better mechanical properties and more intact vasculature structure, facilitating the induction of angiogenesis post‐implantation . Also, the presence of ECM proteins and functional peptides in 3D scaffolds allows recreating environments more physiological for cells, as opposed to traditional 2D culture systems on flat surfaces.…”

“…35 The decellularized tissues were used to seed cells and islets and their in vitro functions were studied as well as the biocompatibility of these bioartificial pancreata was investigated in vivo. [34][35][36][37] Later, rat pancreata were decellularized and a new method was introduced to infuse islets into the ducts, veins and arteries. 38 Decellularized pancreata were seeded with human stem cells, an AR427 acinal cell line, MIN-6 cells and islets, and cell attachment as well as basic cell functions was studied.…”