Quantifying cellular protrusion in alginate capsules with covalently crosslinked shells

Abstract

This work describes viability and distribution of INS-1E beta cells in shell-crosslinked alginate capsules, focussing on cells located near the capsule surface. Capsules were formed by air-shearing alginate suspensions of INS-1E cells into a gelling bath, and coating with poly-l-lysine (PLL) and 50% hydrolysed poly(methylvinylether-alt-maleic anhydride) to form crosslinked networks reinforcing the capsule surfaces. The percentage of cells at the capsule surface were determined using 2D and 3D confocal colocalization mapping. Encapsulated INS-1E cells showed high cell viability and progressive cell clustering out to six weeks. About 30% of cells were initially colocated with the 20 micrometer thick alginate-PLL-PMM50 shell, with 7% of cells protruded at the capsule surfaces, both reflecting random cell distributions. Protruding cells may cause cell-based immune responses, weaken capsules, and potentially result in cell escape from the capsules. The data shown indicate that reinforcing capsules with crosslinked shells may assist in preventing cell exposure and escape.

Keywords:

• Word
• cell encapsulation
• covalently crosslinked shells
• cellular protrusion
• INS-1E cells

Acknowledgements

Dr. Claes Wollheim (University of Geneva, Switzerland) for providing the INS-1E cell line used in this research. The authors would like to acknowledge NSERC for Discovery Grants to both A. Holloway and H. Stöver to fund this research.

Disclosure statement

HDHS and NADB declare financial interests in patents filed on the encapsulation technology. HDHS is involved in a start-up company on cell encapsulation.