Abstract

The technique of immunoisolated transplantation has seen in the last twenty years improvements in biocompatibility, long term stability and methods for avoidance of fibrosis in alginate capsules. However, two major problems are not yet solved: living cellular material that is not centered in the capsule is not properly protected from the hosts’ immune system and the total transplant volume needs to be reduced. To solve these problems, we present a method for applying fully biocompatible alginate multilayers to a barium-alginate core without the use of polycations. We report on the factors that influence layer formation and stability and can therefore provide data for full adjustability of the additional layer. Although known for yeast and plant cells, this technique has not previously been demonstrated with mammalian cells or ultra-high viscous alginates. Viability of murine insulinoma cells was investigated by live-dead staining and live cell imaging, for murine Langerhans’ islets viability and insulin secretion have been measured. No hampering effects of the second alginate layer were found. This multi-layer technique therefore has great potential for clinical and in vitro use and is likely to be central in alginate matrix based immunoisolated cell therapy.

Highlights

  • Encapsulation of cells has a long history in biotechnology especially for the immunoisolated transplantation of endocrine cells or tissues

  • Glucose-stimulated insulin secretion was expressed as stimulation index (SI) and calculated as the ratio of insulin released during exposure to high glucose over the insulin released during low glucose incubation (5.5 mM and 16.6 mM, respectively)

  • An alginate layer was added according to the protocol using different methods for BaSO4 crystal application and different concentrations of alginate

Read more

Summary

Introduction

Encapsulation of cells has a long history in biotechnology especially for the immunoisolated transplantation of endocrine cells or tissues. Alginate is the preferred material for this application because it is biocompatible, polymerizes under gentle conditions and is non toxic in both the polymerized state and in solution. It is an unbranched polymer containing manuronic- (M) and guluronic (G) acid. Capsule formation occurs when droplets of liquid alginate fall into a polymerization bath containing divalent cations, whereas different methods and techniques for droplet formation are yet established [3,6]. Hydrogels made of UHV (ultra high viscous) alginates, characterized by extraordinary high molecular weight, have special properties, which makes them ideal for biomedical applications especially in the field of long term immunoisolation of transplanted cells [4,9,10]. UHV alginate sheets can be very stable and under special conditions withstand pressures of 2 bar [12]

Methods
Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call