Low-adhesive ethylene vinyl alcohol-based packaging to xenogeneic islet encapsulation for type 1 diabetes treatment.

Transplantation of encapsulated porcine islets is proposed to treat type 1 diabetes. However, the envelopment of fibrous tissue and the infiltration of immune cells impair islet function and eventually cause implant failure. It is known that hemodialysis using an ethylene vinyl alcohol (EVOH) membrane results in minor tissue responses. Therefore, we hypothesized that using a low-adhesive EVOH membrane for encapsulation may prevent host cell accumulation and fibrous capsule formation. In this study, rat islets suspended in chitosan gel were encapsulated in bags made from highly porous EVOH membranes, and their in vitro insulin secretion function as well as in vivo performance was evaluated. The results showed that the EVOH bag did not affect islet survival or glucose-stimulated insulin secretion. Whereas naked islets were dysfunctional after 7 days of culture in vitro, islets within the EVOH bag produced insulin continuously for 30 days. Streptozotocin-induced diabetic mice were given islets-chitosan gel-EVOH implants intraperitoneally (650-800 islets equivalent) and exhibited lower blood glucose levels and regained body weight during a 4-week observation period. The transplanted mice had higher levels of serum insulin and C-peptide, with an improved blood glucose disappearance rate. Retrieved implants had minor tissue adhesion, and histology showed a limited number of mononuclear cells and fibroblasts surrounding the implants. No invasion of host cells into the EVOH bags was noticed, and the encapsulated islets were intact and positive for insulin-glucagon immunostaining. In conclusion, an EVOH bag can protect encapsulated islets, limit fibrous capsule formation, and extend graft function.

A multiple-funnels cell culture insert for the scale-up production of uniform cell spheroids.

INTRODUCTION: Formation of cell spheres is an important procedure in biomedical research. A large number of high-quality cell spheres of uniform size and shape are required for basic studies and therapeutic applications. Conventional approaches, including the hanging drop method and suspension culture, are used for cell sphere production. However, these methods are time consuming, cell spheres cannot be harvested easily, and it is difficult to control the size and geometry of cell spheres. To resolve these problems, a novel multiple-funnel cell culture insert was designed for size controlling, easy harvesting, and scale-up production of cell spheres. METHODS: The culture substrate has 680 micro-funnels with a 1-mm width top, 0.89 mm depth, and 0.5 mm square bottom. Mouse embryonic stem cells were used to test the newly developed device. The seeded embryonic stem cells settled at the downward medium surface toward the bottom opening and aggregated as embryoid bodies (EBs). For cell sphere harvest, the bottom of the culture insert was put in contact with the medium surface in another culture dish, and the medium in the device flowed down with cell spheres by hydrostatic pressure. RESULTS: Compact cell spheres with uniform size and shape were collected easily. The diameter of the spheres could be controlled by adjusting the seeding cell density. Spontaneous neural differentiation (nestin and Tju1) and retinoic acid-induced endodermal differentiation (Pdx-1 and insulin I) were improved in the EBs produced using the new insert compared to those in EBs produced by suspension culture. CONCLUSIONS: This novel cell culture insert shall improve future studies of cell spheres and benefit clinical applications of cell therapy.

Effect of swelling of poly(vinyl alcohol) layers on complement activation.

Polymers carrying hydroxyl groups have the potential ability to activate the complement system when in contact with blood. However, the effects of their surface structure on complement activation are still not fully understood. In this study, we examined complement activation by poly(vinyl alcohol) (PVA) layers formed on a gold surface modified with aldehyde groups. The complement system was strongly activated by a PVA surface with a dry thickness of 2.9 nm, while it was poorly activated by a PVA surface with a dry thickness of 7.4 nm. Annealing of the latter for 2 h at 150 degrees C converted the surface into a complement activating surface. The difference in complement activation between PVA layers was associated with the water content of PVA layers. These results suggest that complement activation by hydrated polymers highly depends on the water content of the polymer layers.

Complement activation by polymers carrying hydroxyl groups.

Hydrogels of polymers carrying surface hydroxyl groups strongly activate the complement system through the alternative pathway, although it has also been reported that solutions of polymers do not. To address these curious, inconsistent results, we examined the effect of polymer states, either immobilized on a surface or soluble in serum, on the complement activation using a surface plasmon resonance apparatus and enzyme-linked immunosorbent assay. We clearly showed that dextran- and poly(vinyl alcohol)-immobilized surfaces strongly activated the complement system but that soluble polymers could not, even when the amounts of the soluble polymers added to serum were 4-2000 times higher than those on the polymer-immobilized surfaces.